Acylated active agents and methods of their use for the treatment of autoimmune disorders

ABSTRACT

Disclosed herein are acylated active agents and methods of their use, e.g., for modulating an autoimmunity marker or for treating an autoimmune disorder.

FIELD OF THE INVENTION

The invention relates compounds and methods of their medicinal use.

BACKGROUND

Autoimmune disorders are the result of one's own immune systemincorrectly attacking one's own tissue. Inflammatory bowel disease is agroup of autoimmune disorders (e.g., Crohn's disease and ulcerativecolitis). It is believed that inflammatory bowel disease relates to ahost immune response toward the resident microbiota. The currentstandard of care for inflammatory bowel disease includes steroid andimmunosuppressant therapies, which often produce undesirable sideeffects and can carry substantial risks of comorbidity development.

There is a need for new approaches to the treatment of autoimmunedisorders.

SUMMARY OF THE INVENTION

In general, the invention provides acylated active agents (e.g.,acylated catechin polyphenols, acylated carotenoids, acylatedmesalamines, acylated sugars, acylated shikimic acids, acylated ellagicacid, acylated ellagic acid analogue, and acylated hydroxybenzoicacids), active agent combinations (e.g., a first agent that is catechinpolyphenol, carotenoid, mesalamine, shikimic acid, or hydroxybenzoicacid and a second agent that is a fatty acid), compositions containingthem (e.g., as unit dosage forms), and methods for modulating anautoimmunity marker in a subject or of treating an autoimmunity disorderin a subject. In some embodiments,

In one aspect, the invention provides a method of modulating anautoimmunity marker in a subject in need thereof by administering to thesubject an effective amount of an active agent. In a related aspect, theinvention provides a method of treating an autoimmunity disorder in asubject in need thereof by administering to the subject an effectiveamount of an active agent.

In some embodiments, the autoimmunity marker is for an inflammatorybowel disease, Addison's disease, alopecia areata, ankylosingspondylitis, antiphospholipid syndrome, hemolytic anemia, autoimmunehepatitis, Behcet's disease, Berger's disease, bullous pemphigoid,cardiomyopathy, celiac sprue, chronic fatigue immune dysfunctionsyndrome (CFIDS), chronic inflammatory demyelinating polyneuropathy,Churg-Strauss syndrome, cicatricial pemphigoid, cold agglutinin disease,type 1 diabetes, discoid lupus, essential mixed cryoglobulinemia,Graves' disease, Guillain-Barré syndrome, Hashimoto's thyroiditis,hypothyroidism, autoimmune lymphoproliferative syndrome (ALPS),idiopathic pulmonary fibrosis, idiopathic thrombocytopenia purpura(ITP), juvenile arthritis, lichen planus, lupus erythematosus, Meniere'sdisease, mixed connective tissue disease, multiple sclerosis, myastheniagravis, pemphigus vulgaris, pernicious anemia, polychondritis,autoimmune polyglandular syndromes, polymyalgia rheumatica,polymyositis, dermatomyositis, primary agammaglobulinemia, primarybiliary cirrhosis, psoriasis, psoriatic arthritis, Raynaud's phenomenon,Reiter's syndrome, rheumatic fever, rheumatoid arthritis, sarcoidosis,scleroderma, Sjögren's syndrome, stiff-man syndrome, Takayasu arteritis,giant cell arteritis, ulcerative colitis, uveitis, vasculitis, orgranulomatosis with polyangiitis.

In certain embodiments, the autoimmunity marker is for an inflammatorybowel disease (e.g., ulcerative colitis or Crohn's disease). Inparticular embodiments, the subject suffers from the autoimmunedisorder. In some embodiments, the autoimmune disorder is rheumatoidarthritis.

In further embodiments, the autoimmune disorder is an inflammatory boweldisease, Addison's disease, alopecia areata, ankylosing spondylitis,antiphospholipid syndrome, hemolytic anemia, autoimmune hepatitis,Behcet's disease, Berger's disease, bullous pemphigoid, cardiomyopathy,celiac sprue, chronic fatigue immune dysfunction syndrome (CFIDS),chronic inflammatory demyelinating polyneuropathy, Churg-Strausssyndrome, cicatricial pemphigoid, cold agglutinin disease, type 1diabetes, discoid lupus, essential mixed cryoglobulinemia, Graves'disease, Guillain-Barré syndrome, Hashimoto's thyroiditis,hypothyroidism, autoimmune lymphoproliferative syndrome (ALPS),idiopathic pulmonary fibrosis, idiopathic thrombocytopenia purpura(ITP), juvenile arthritis, lichen planus, lupus erythematosus, Meniere'sdisease, mixed connective tissue disease, multiple sclerosis, myastheniagravis, pemphigus vulgaris, pernicious anemia, polychondritis,autoimmune polyglandular syndromes, polymyalgia rheumatica,polymyositis, dermatomyositis, primary agammaglobulinemia, primarybiliary cirrhosis, psoriasis, psoriatic arthritis, Raynaud's phenomenon,Reiter's syndrome, rheumatic fever, rheumatoid arthritis, sarcoidosis,scleroderma, Sjögren's syndrome, stiff-man syndrome, Takayasu arteritis,giant cell arteritis, ulcerative colitis, uveitis, vasculitis, orgranulomatosis with polyangiitis.

In some embodiments, the autoimmune disorder is rheumatoid arthritis.

In yet further embodiments, the autoimmune disorder is an inflammatorybowel disease (e.g., ulcerative colitis or Crohn's disease). Inparticular embodiments, the vasculitis is polyarteritis nodosa.

In some embodiments, a CYP1A1 mRNA level, intestinal motility, CD4⁺CD25⁺Treg cell count, short chain fatty acids level, or mucus secretion isincreased following the step of administering. In further embodiments, aCYP1A1 mRNA level is increased following the administration of theacylated active agent to the subject. In certain embodiments, abdominalpain, gastrointestinal inflammation, gastrointestinal permeability,gastrointestinal bleeding, intestinal motility, or frequency of bowelmovements is reduced following the step of administering. In particularembodiments, an interleukin-8 (IL-8) level, macrophage inflammatoryprotein 1α (MIP-1α) level, macrophage inflammatory protein 1β (MIP-1β)level, NFκB level, inducible nitric oxide synthase (iNOS) level, matrixmetallopeptidase 9 (MMP9) level, interferon γ (IFNγ) level,interleukin-17 (IL17) level, intercellular adhesion molecule (ICAM)level, CXCL13 level, 8-iso-prostaglandin F_(2α) (8-iso-PGF2α) level IgAlevel, calprotectin level, lipocalin-2 level, or indoxyl sulfate levelis reduced following the step of administering. In some embodiments, aninterleukin-8 (IL-8) level, macrophage inflammatory protein 1α (MIP-1α)level, or macrophage inflammatory protein 1β (MIP-1β) level is reducedfollowing the administration of the acylated active agent to thesubject. In further embodiments, the T_(h)1 cell count is modulatedfollowing the step of administering.

In some embodiments, the active agent is an acylated catechin polyphenolor acylated mesalamine. In particular embodiments, following oraladministration to the subject, the active agent is hydrolyzable in thegastrointestinal tract of the subject. In certain embodiments, theactive agent releases at least one fatty acid. In further embodiments,the active agent is administered to the subject orally.

In certain embodiments, the active agent is acylated mesalamine. In someembodiments, the active agent is an acylated catechin polyphenol.

In yet further embodiments, the active agent is an acylated hydroxybenzoic acid. In still further embodiments, the acylated hydroxybenzoicacid includes a core selected from the group consisting of salicylicacid and gallic acid.

In certain embodiments, the active agent is an acylated sugar. In someembodiments, the acylated sugar includes a monosaccharide core (e.g.,xylose, arabinose, rhamnose, fucose, glucosamine, tagatose, or ribose).In further embodiments, the monosaccharide core is xylose. In yetfurther embodiments, the monosaccharide core is arabinose. In stillfurther embodiments, the monosaccharide core is rhamnose. In otherembodiments, the monosaccharide core is fucose. In yet otherembodiments, the monosaccharide core is glucosamine. In still otherembodiments, the monosaccharide core is tagatose. In some embodiments,the monosaccharide core is ribose. In certain embodiments, the acylatedsugar comprises a sugar acid core. In particular embodiments, the sugaracid core is a uronic acid (e.g., glucuronic acid).

In particular embodiments, the active agent is an acylated shikimicacid. In some embodiments, an acylated shikimic acid is of the followingstructure:

or a salt thereof, where

each R¹ is independently H, acyl, alkyl, a group containing a fattyacid, a group containing a ketone body or pre-ketone body, or a groupcontaining an amino acid metabolite; and

R² is H, alkyl, a group containing a fatty acid, a group containing aketone body or pre-ketone body, or a group containing an amino acidmetabolite;

provided that the compound includes at least one group containing afatty acid, group containing a ketone body or pre-ketone body, or groupcontaining an amino acid metabolite.

In certain embodiments, at least one R¹ is a group containing a fattyacid. In particular embodiments, at least one R¹ is a group containing aketone body or pre-ketone body. In some embodiments, at least one R¹ isa group containing an amino acid metabolite. In further embodiments, R²is H.

In other embodiments, the acylated active agent includes a groupcontaining a fatty acid. In yet other embodiments, the group containinga fatty acid is a monosaccharide (e.g., arabinose, xylose, fructose,galactose, glucose, glucosinolate, ribose, tagatose, fucose, andrhamnose), sugar alcohol, or sugar acid having one or more hydroxylgroups substituted with a fatty acid acyl). In still other embodiments,the monosaccharide is L-arabinose, D-xylose, fructose, galactose,D-glucose, glucosinolate, D-ribose, D-tagatose, L-fucose, or L-rhamnose(e.g., the monosaccharide is D-xylose). In further embodiments, thegroup containing a fatty acid is a fatty acid acyl. In yet furtherembodiments, the fatty acid is a short chain fatty acid (e.g., acetyl,propionyl, or butyryl). In still further embodiments, the short chainfatty acid is acetyl. In particular embodiments, the short chain fattyacid is butyryl. In certain embodiments, the fatty acid is a mediumchain fatty acid (e.g., octanoyl).

In some embodiments, the acylated active agent includes a groupcontaining a ketone body or pre-ketone body. In certain embodiments, thegroup containing a ketone body or pre-ketone body is a monosaccharide,sugar alcohol, or sugar acid having one or more hydroxyl groupssubstituted with a ketone body acyl or pre-ketone body acyl. Inparticular embodiments, the group containing a ketone body or pre-ketonebody includes at least one ketone body. In further embodiments, thegroup containing a ketone body or pre-ketone body includes at least onepre-ketone body. In yet further embodiments, the ketone body isβ-hydroxybutyric acid. In still further embodiments, the pre-ketone bodyis 1,3-butanediol. In still further embodiments, the group containing aketone body or pre-ketone body is a ketone body acyl or a pre-ketonebody acyl.

In certain embodiments, the acylated active agent includes a groupcontaining an amino acid metabolite. In particular embodiments, thegroup containing an amino acid metabolite is a monosaccharide, sugaralcohol, or sugar acid having one or more hydroxyl groups substitutedwith an amino acid metabolite acyl. In some embodiments, the groupcontaining an amino acid metabolite is an amino acid metabolite acyl. Infurther embodiments, the amino acid metabolite is indole-3-acetic acid,indole-3-acrylic acid, or indole-3-pyruvic acid.

In still further embodiments, the acylated catechin polyphenol is acompound of formula (I):

or a pharmaceutically acceptable salt thereof,

wherein

is a single carbon-carbon bond or double carbon-carbon bond;

Q is —CH₂— or —C(O)—;

each R¹ and each R³ is independently H, halogen, —OR^(A), phosphate, orsulfate;

R² is H or —OR^(A);

each R^(A) is independently H, optionally substituted alkyl, amonosaccharide, a sugar acid, a group containing a fatty acid, a groupcontaining a ketone body or pre-ketone body, a group containing an aminoacid metabolite, or benzoyl optionally substituted with 1, 2, 3, or 4substituents independently selected from the group consisting of H,hydroxy, halogen, a group containing a fatty acid, a group containing aketone body or pre-ketone body, a group containing an amino acidmetabolite, an optionally substituted alkyl, an optionally substitutedalkoxy, a monosaccharide, a sugar acid, phosphate, and sulfate;

each of n and m is independently 0, 1, 2, 3, or 4.

In particular embodiments, the compound of formula (I) includes at leastone group containing a fatty acid, group containing a ketone body orpre-ketone body, or group containing an amino acid metabolite; and atleast one group containing a fatty acid, when present, is amonosaccharide having one, two, three, or four hydroxyls substitutedwith fatty acid acyls.

In some embodiments, at least one R¹ is —OR^(A), in which R^(A) is agroup containing a fatty acid, group containing a ketone body orpre-ketone body, or group containing an amino acid metabolite.

In certain embodiments, the acylated catechin polyphenol is a compoundis of formula (I-a):

In particular embodiments, the acylated catechin polyphenol is acompound is of formula (I-b):

In further embodiments, the acylated catechin polyphenol is a compoundis of formula (I-c):

In yet further embodiments, the acylated catechin polyphenol is acompound is of formula (I-d):

In certain embodiments, the acylated catechin polyphenol is a compoundof formula (I-f):

In still further embodiments, n is 2. In certain embodiments, m is 1. Inparticular embodiments, m is 2. In some embodiments, m is 3. Inparticular embodiments, each R¹ is independently-OR^(A). In certainembodiments, each R³ is independently H or —OR^(A). In furtherembodiments, R² is H or —OR^(A). In yet further embodiments, each R^(A)is independently H, optionally substituted alkyl, a group containing afatty acid, a group containing a ketone body or pre-ketone body, or agroup containing an amino acid metabolite.

In other embodiments, the acylated catechin polyphenol is a compound isof formula (I-e):

or a pharmaceutically acceptable salt thereof,

wherein each of R^(1A) and R^(1B) is independently as defined for R¹;and each of R^(3A), R^(3B), and R^(3C) is independently as defined forR³.

In yet other embodiments, each of R^(1A) and R^(1B) is independently—OR^(A). In still other embodiments, each of R^(3A), R^(3B), and R^(3C)is independently H, halogen, or —OR^(A). In some embodiments, R² is agroup of formula:

wherein p is 1, 2, 3, or 4, and each R⁴ is independently selected fromthe group consisting of H, hydroxy, halogen, a group containing a fattyacid, a group containing a ketone body or pre-ketone body, a groupcontaining an amino acid metabolite, an optionally substituted alkyl, anoptionally substituted alkoxy, a monosaccharide, a sugar acid,phosphate, and sulfate.

In certain embodiments, p is 3. In particular embodiments, each R⁴ isindependently H, hydroxy, halogen, a group containing a fatty acid, agroup containing a ketone body or pre-ketone body, a group containing anamino acid metabolite, or an optionally substituted alkoxy. In certainembodiments, R² is a group of formula:

each of R^(4A), R^(4B), and R^(4C) is as defined for R⁴.

In further embodiments, each of R^(4A), R^(4B), and R^(4C) isindependently H, hydroxy, halogen, a group containing a fatty acid, agroup containing a ketone body or pre-ketone body, a group containing anamino acid metabolite, or an optionally substituted alkoxy. In yetfurther embodiments, each R^(A) is independently H, optionallysubstituted alkyl, fatty acid acyl, or optionally acylatedmonosaccharide.

In still further embodiments, the acylated active agent includes atleast one fatty acid acyl (e.g., a short chain fatty acid acyl). In someembodiments, the short chain fatty acid acyl is acetyl, propionyl, orbutyryl. In certain embodiments, the short chain fatty acid acyl isacetyl. In particular embodiments, the short chain fatty acid acyl isbutyryl.

In another aspect, the invention provides, a composition (e.g., apharmaceutical composition, nutraceutical composition, food product,food additive, or dietary supplement) including an active agent. In someembodiments, the composition is provided in a unit dosage form. In otherembodiments, the active agent is an acylated catechin polyphenol,acylated carotenoid, acylated mesalamine, acylated sugar, acylatedshikimic acid, acylated ellagic acid, acylated ellagic acid analogue,and acylated hydroxybenzoic acid.

In still other embodiments, the unit dosage form contains at least 0.5 g(e.g., at least 0.7 g, at least 1 g, or at least 2 g) of the activeagent. In certain embodiments, the unit dosage form contains 10 g orless (e.g., 9 g or less, 8 g or less, 7 g or less, 6 g or less, 5 g orless) of the active agent. In particular embodiments, the unit dosageform contains 0.5-10 g (e.g., 0.7-10 g, 1-10 g, 2-10 g, 0.5-9 g, 0.7-9g, 1-9 g, 2-9 g, 0.5-8 g, 0.7-8 g, 1-8 g, 2-8 g, 0.5-7 g, 0.7-7 g, 1-7g, 2-7 g, 0.5-6 g, 0.7-6 g, 1-6 g, 2-6 g, 0.5-5 g, 0.7-5 g, 1-10 g, or2-5 g) of the active agent.

In some embodiments, the unit dosage form is a pharmaceutical unitdosage form. In further embodiments, the unit dosage form is anutraceutical dosage form. In yet further embodiments, the unit dosageform is a serving of a food product.

In still further embodiments, the active agent is the acylated catechinpolyphenol. In certain embodiments, the active agent is an acylatedmesalamine. In yet further embodiments, the active agent is an acylatedhydroxybenzoic acid. In still further embodiments, the active agent isan acylated sugar. In particular embodiments, the active agent is anacylated shikimic acid. In some embodiments, the active agent is anacylated ellagic acid. In certain embodiments, the active agent is anacylated ellagic acid analogue (e.g., an analogue including urolithin Ccore). In particular embodiments, the active agent is an acylatedcarotenoid.

In other embodiments, the acylated active agent includes a groupcontaining a fatty acid. In yet other embodiments, the group containinga fatty acid is a monosaccharide (e.g., arabinose, xylose, fructose,galactose, glucose, glucosinolate, ribose, tagatose, fucose, andrhamnose), sugar alcohol, or sugar acid having one or more hydroxylgroups substituted with a fatty acid acyl). In still other embodiments,the monosaccharide is L-arabinose, D-xylose, fructose, galactose,D-glucose, glucosinolate, D-ribose, D-tagatose, L-fucose, or L-rhamnose(e.g., the monosaccharide is D-xylose). In further embodiments, thegroup containing a fatty acid is a fatty acid acyl. In yet furtherembodiments, the fatty acid is a short chain fatty acid (e.g., acetyl,propionyl, or butyryl). In still further embodiments, the short chainfatty acid is acetyl. In particular embodiments, the short chain fattyacid is butyryl. In certain embodiments, the fatty acid is a mediumchain fatty acid (e.g., octanoyl).

In other embodiments, the acylated active agent includes a groupcontaining a ketone body or pre-ketone body. In yet other embodiments,the group containing a ketone body or pre-ketone body is amonosaccharide (e.g., arabinose, xylose, fructose, galactose, glucose,glucosinolate, ribose, tagatose, fucose, and rhamnose), sugar alcohol,or sugar acid having one or more hydroxyl groups substituted with aketone body acyl or pre-ketone body acyl). In still other embodiments,the monosaccharide is L-arabinose, D-xylose, fructose, galactose,D-glucose, glucosinolate, D-ribose, D-tagatose, L-fucose, or L-rhamnose(e.g., the monosaccharide is D-xylose). In further embodiments, thegroup containing a ketone body or pre-ketone body is a ketone body acylor pre-ketone body acyl.

In other embodiments, the acylated active agent includes a groupcontaining an amino acid metabolite. In yet other embodiments, the groupcontaining an amino acid metabolite is a monosaccharide (e.g.,arabinose, xylose, fructose, galactose, glucose, glucosinolate, ribose,tagatose, fucose, and rhamnose), sugar alcohol, or sugar acid having oneor more hydroxyl groups substituted with an amino acid metabolite acyl).In still other embodiments, the monosaccharide is L-arabinose, D-xylose,fructose, galactose, D-glucose, glucosinolate, D-ribose, D-tagatose,L-fucose, orL-rhamnose (e.g., the monosaccharide is D-xylose). Infurther embodiments, the group containing an amino acid metabolite is anamino acid metabolite acyl.

In still further embodiments, the acylated catechin polyphenol is acompound of formula (I):

or a pharmaceutically acceptable salt thereof,

wherein

is a single carbon-carbon bond or double carbon-carbon bond;

Q is —CH₂— or —C(O)—;

each R¹ and each R³ is independently H, halogen, —OR^(A), phosphate, orsulfate;

R² is H or —OR^(A);

each R^(A) is independently H, optionally substituted alkyl, amonosaccharide, a sugar acid, a group containing a fatty acid, a groupcontaining a ketone body or pre-ketone body, a group containing an aminoacid metabolite acyl, or benzoyl optionally substituted with 1, 2, 3, or4 substituents independently selected from the group consisting of H,hydroxy, halogen, a group containing a fatty acid, a group containing aketone body or pre-ketone body, a group containing an amino acidmetabolite acyl, an optionally substituted alkyl, an optionallysubstituted alkoxy, a monosaccharide, a sugar acid, phosphate, andsulfate;

each of n and m is independently 0, 1, 2, 3, or 4.

In particular embodiments, the compound of formula (I) includes at leastone group containing a fatty acid, group containing a ketone body orpre-ketone body, or group containing an amino acid metabolite; and atleast one group containing a fatty acid, when present, is amonosaccharide having one, two, three, or four hydroxyls substitutedwith fatty acid acyls.

In some embodiments, at least one R¹ is —OR^(A), in which R^(A) is agroup containing a fatty acid, group containing a ketone body orpre-ketone body, or group containing an amino acid metabolite acyl.

In certain embodiments, the acylated catechin polyphenol is a compoundis of formula (I-a):

In particular embodiments, the acylated catechin polyphenol is acompound is of formula (I-b):

In further embodiments, the acylated catechin polyphenol is a compoundis of formula (I-c):

In yet further embodiments, the acylated catechin polyphenol is acompound is of formula (I-d):

In certain embodiments, the acylated catechin polyphenol is a compoundof formula (I-f):

In still further embodiments, n is 2. In certain embodiments, m is 1. Inparticular embodiments, m is 2. In some embodiments, m is 3. Inparticular embodiments, each R¹ is independently-OR^(A). In certainembodiments, each R³ is independently H or —OR^(A). In furtherembodiments, R² is H or —OR^(A). In yet further embodiments, each R^(A)is independently H, optionally substituted alkyl, a group containing afatty acid, a group containing a ketone body or pre-ketone body, or agroup containing an amino acid metabolite. In some embodiments, at leastone R^(A) is a group containing a fatty acid. In certain embodiments, atleast one R^(A) is a group containing an amino acid metabolite. Inparticular embodiments, at least one R^(A) is a group containing aketone body or pre-ketone body. Preferably, the compound of formula(I-f) includes quercetin core.

In other embodiments, the acylated catechin polyphenol is a compound isof formula (I-e):

or a pharmaceutically acceptable salt thereof,

wherein each of R^(1A) and R^(1B) is independently as defined for R¹;and each of R^(3A), R^(3B), and R^(3C) is independently as defined forR³.

In yet other embodiments, each of R^(1A) and R^(1B) is independently—OR^(A). In still other embodiments, each of R^(3A), R^(3B), and R^(3C)is independently H, halogen, or —OR^(A). In some embodiments, R² is agroup of formula:

wherein p is 1, 2, 3, or 4, and each R⁴ is independently selected fromthe group consisting of H, hydroxy, halogen, a group containing a fattyacid, group containing a ketone body or pre-ketone body, groupcontaining an amino acid metabolite, an optionally substituted alkyl, anoptionally substituted alkoxy, a monosaccharide, a sugar acid,phosphate, and sulfate.

In certain embodiments, p is 3. In particular embodiments, each R⁴ isindependently H, hydroxy, halogen, a group containing a fatty acid,group containing a ketone body or pre-ketone body, group containing anamino acid metabolite acyl, or an optionally substituted alkoxy. Incertain embodiments, R² is a group of formula:

and

each of R^(4A), R^(4B), and R^(4C) is as defined for R⁴.

In further embodiments, each of R^(4A), R^(4B), and R^(4C) isindependently H, hydroxy, halogen, a group containing a fatty acid,group containing a ketone body or pre-ketone body, group containing anamino acid metabolite acyl, or an optionally substituted alkoxy. In yetfurther embodiments, each R^(A) is independently H, optionallysubstituted alkyl, fatty acid acyl, or optionally acylatedmonosaccharide. In still further embodiments, the acylated catechinpolyphenol includes at least one fatty acid acyl (e.g., a short chainfatty acid acyl). In some embodiments, the short chain fatty acid acylis acetyl, propionyl, or butyryl. In certain embodiments, the shortchain fatty acid acyl is acetyl. In particular embodiments, the shortchain fatty acid acyl is butyryl.

Definitions

The term “acyl,” as used herein, represents a chemical substituent offormula —C(O)—R, where R is alkyl, alkenyl, aryl, arylalkyl, cycloalkyl,heterocyclyl, heterocyclyl alkyl, heteroaryl, or heteroaryl alkyl. Anoptionally substituted acyl is an acyl that is optionally substituted asdescribed herein for each group R. Non-limiting examples of acyl includefatty acid acyls (e.g., short chain fatty acid acyls (e.g., acetyl)),amino acid metabolite acyl, ketone body acyl, pre-ketone body acyl, andbenzoyl.

The term “acylated active agent,” as used herein, represents a compoundincluding two or more agents linked through ester bond(s), amidebond(s), carbonate linker(s), carbamate linker(s), and/or glycosidicbond(s). Non-limiting examples of acylated active agents include anacylated catechin polyphenol and acylated mesalamine.

The term “acylated catechin polyphenol,” as used herein, represents asubstituted compound having the core of formula (A):

or a multimer thereof, or a salt thereof,

where the substituents are independently selected from the groupconsisting of —OR^(A), —OCOO—R^(A), —NHR^(B), oxo, halogen, optionallysubstituted C₁₋₂₀ alkyl, optionally substituted C₂₋₂₀ alkenyl,optionally substituted thioalkyl, optionally substituted alkylsulfonyl,optionally substituted alkylsulfenyl, optionally substitutedalkylsulfinyl, optionally substituted thioaryl, optionally substitutedaryl thioalkyl, optionally substituted thioalkenyl, dialkylamino,sulfate, phosphate, ascorbic acid, optionally substituted heterocyclyl,nitro, amino acids, C₁₋₆ esters of amino acids, optionally acylatedmonosaccharide, and optionally acylated sugar acid, where each R^(A) isindependently H, optionally substituted alkyl, a group containing afatty acid, a group containing a ketone body or pre-ketone body, orbenzoyl optionally substituted with one, two, three, or foursubstituents independently selected from the group consisting of H,hydroxyl, halogen, a group containing a fatty acid, a group containing aketone body or pre-ketone body, optionally substituted alkoxy, andoptionally substituted alkyl, and where R^(B) is independently H oroptionally substituted alkyl;

where the carbon-carbon bond connecting carbon 2 and carbon 3 in formula(A) is a single bond or a double bond;

where the multimer includes a total of 2 or 3 cores of formula (A), eachcore substituted independently as described above; and

where two vicinal centers in core (A) may be further substituted with agroup —(O)_(q)-L¹-L²-, where q is 0 or 1, L¹ is optionally substitutedalkylene, optionally substituted alkenylene, or optionally substitutedheterocyclylene; and L² is a covalent bond, optionally substitutedheterocyclylene, or optionally substituted cycloalkylene;

provided that at least one of positions 5, 6, 7, and 8 is —OR^(A), whereR^(A) is a group containing a fatty acid, a group containing a ketonebody or pre-ketone body, or a benzoyl optionally substituted with one,two, three, or four substituents independently selected from the groupconsisting of H, hydroxyl, a halogen, a group containing a fatty acid, agroup containing a ketone body or pre-ketone body, an optionallysubstituted alkoxy, and an optionally substituted alkyl; and providedthat the substituted compound includes at least one group containing afatty acid, group containing a ketone body or pre-ketone body, or groupcontaining an amino acid metabolite acyl. Preferably, the acylatedcatechin polyphenol includes quercetin core.

The term “acylated carotenoid,” as used herein, represents a carotenoid,in which at least one hydroxyl group is replaced with a substituent —OR,where each R is independently selected from the group consisting of anacyl, alkyl, a group containing a fatty acid, a group containing aketone body or pre-ketone body, and a group containing an amino acidmetabolite, provided that at least one R is a group containing a fattyacid, a group containing a ketone body or pre-ketone body, or a groupcontaining an amino acid metabolite. Non-limiting examples of anacylated carotenoid include astaxanthin having one or both hydroxylgroups independently substituted with an acyl, alkyl, group containing afatty acid, or group containing a ketone body or pre-ketone body,provided that at least one hydroxyl is substituted with a groupcontaining a fatty acid, a group containing a ketone body or pre-ketonebody, or a group containing an amino acid metabolite.

The term “acylated catechin polyphenol” also refers to a compound offormula (I):

or a pharmaceutically acceptable salt thereof,where

is a single carbon-carbon bond or double carbon-carbon bond;

Q is —CH₂— or —C(O)—;

each R¹ and each R³ is independently H, halogen, —OR^(A), phosphate, orsulfate;

R² is H or —OR^(A);

each R^(A) is independently H, optionally substituted alkyl, amonosaccharide, a sugar acid, a group containing a fatty acid, a groupcontaining a ketone body or pre-ketone body, a group containing an aminoacid metabolite, or benzoyl optionally substituted with 1, 2, 3, or 4substituents independently selected from the group consisting of H,hydroxy, halogen, a group containing a fatty acid, a group containing aketone body or pre-ketone body, or a group containing an amino acidmetabolite an optionally substituted alkyl, an optionally substitutedalkoxy, a monosaccharide, a sugar acid, phosphate, and sulfate; and

each of n and m is independently 0, 1, 2, 3, or 4.

The term “acylated hydroxybenzoic acid,” as used herein, represents acompound of formula:

or a salt thereof,

where

n is 1, 2, or 3;

each R¹ is independently H, acyl, alkyl, a group containing a fattyacid, a group containing a ketone body or pre-ketone body, or a groupcontaining an amino acid metabolite; and

R² is H, alkyl, a group containing a fatty acid, a group containing aketone body or pre-ketone body, or a group containing an amino acidmetabolite;

provided that the compound includes at least one group containing afatty acid, group containing a ketone body or pre-ketone body, or groupcontaining an amino acid metabolite.

Non-limiting examples of acylated hydroxybenzoic acids include salicylicacid, in which the phenolic hydroxyl is substituted with a groupcontaining a fatty acid, group containing a ketone body or pre-ketonebody, or group containing an amino acid metabolite; and gallic acid, inwhich one, two, or three phenolic hydroxyls are independentlysubstituted with groups containing a fatty acid, group containing aketone body or pre-ketone body, or group containing an amino acidmetabolite.

The term “acylated mesalamine,” as used herein, represents a mesalamine,in which, one H in one or more of —NH₂, —OH, or —COOH is replaced withan acyl or a group containing a fatty acid, group containing a ketonebody or pre-ketone body, or group containing an amino acid metabolite;provided that acylated mesalamine contains at least one group containinga fatty acid, group containing a ketone body or pre-ketone body, orgroup containing an amino acid metabolite. In some embodiments, acylatedmesalamine is a compound of formula (II):

where

R¹ is H, alkyl, acyl, a group containing a fatty acid, a groupcontaining a ketone body or pre-ketone body, or a group containing anamino acid metabolite;

R² is H, alkyl, a group containing a fatty acid, a group containing aketone body or pre-ketone body, or a group containing an amino acidmetabolite; and

each R³ is independently H, alkyl, acyl, a group containing a fattyacid, a group containing a ketone body or pre-ketone body, or a groupcontaining an amino acid metabolite; or both R² groups combine to form:

The acylated mesalamine includes at least one group containing a fattyacid, group containing a ketone body or pre-ketone body, or groupcontaining an amino acid metabolite.

The term “acylated shikimic acid,” as used herein, represents a compoundof formula:

or a salt thereof,

where

each R¹ is independently H, acyl, alkyl, a group containing a fattyacid, a group containing a ketone body or pre-ketone body, or a groupcontaining an amino acid metabolite; and

R² is H, alkyl, a group containing a fatty acid, a group containing aketone body or pre-ketone body, or a group containing an amino acidmetabolite;

provided that the compound includes at least one group containing afatty acid, group containing a ketone body or pre-ketone body, or groupcontaining an amino acid metabolite.

The term “acylated sugar,” as used herein, represents a monosaccharidehaving one or more hydroxyls substituted with alkyl, acyl, a groupcontaining a fatty acid, a group containing a ketone body or pre-ketonebody, or a group containing an amino acid metabolite. The monosaccharideis present in the pyranose or furanose form. Preferably, themonosaccharide is present in the pyranose form. The monosaccharide maybe an aldose or ketose. Non-limiting examples of monosaccharides arearabinose, xylose, fructose, galactose, glucose, ribose, tagatose,fucose, and rhamnose. In some embodiments, the monosaccharide isL-arabinose, D-xylose, fructose, galactose, D-glucose, D-ribose,D-tagatose, L-fucose, or L-rhamnose. Preferably, the monosaccharide isxylose, arabinose, rhamnose, fucose, glucosamine, or tagatose. Themonosaccharide may include an anomeric carbon bonded to —OR, where R isH, alkyl, acyl, a group containing a fatty acid, a group containing aketone body or pre-ketone body, or a group containing an amino acidmetabolite. Preferably, R is alkyl or a group containing a fatty acid.

The term “acylated ellagic acid,” as used herein, represents compoundsof the following structures:

or a salt thereof,

where each R^(A) is independently H, alkyl, acyl, a group containing afatty acid, a group containing a ketone body or pre-ketone body, or agroup containing an amino acid metabolite; and each R^(B) isindependently H, alkyl, a group containing a fatty acid, a groupcontaining a ketone body or pre-ketone body, or a group containing anamino acid metabolite; provided that at least one R^(A) and/or at leastone R^(B), when present, is a group containing a fatty acid, a groupcontaining a ketone body or pre-ketone body, or a group containing anamino acid metabolite.

The term “acylated ellagic acid analogue,” as used herein, representscompounds of the following structure:

or a salt thereof,where

each of R², R³, and R⁴ is independently H or —OR^(A);

R⁶ is H or —(CO)—R^(5B);

R^(1A) is H or —OR^(A), and R^(5A) is —OH or —OR^(B); or R^(1A) andR^(5A) combine to form —O—;

R^(1B) is H or —OR^(A), and R^(5B) is absent, —OH, or —OR^(B); or R^(1B)and R^(5B) combine to form —O—;

each R^(A) is independently H, O-protecting group, alkyl, acyl, a groupcontaining a fatty acid, a group containing a ketone body or pre-ketonebody, or a group containing an amino acid metabolite;

each R^(B) is independently H, O-protecting group, alkyl, a groupcontaining a fatty acid, a group containing a ketone body or pre-ketonebody, or a group containing an amino acid metabolite;

provided that at least one R^(A) and/or at least one R^(B) is a groupcontaining a fatty acid, a group containing a ketone body or pre-ketonebody, or a group containing an amino acid metabolite.

The term “acyloxy,” as used herein, represents a chemical substituent offormula —OR, where R is acyl. An optionally substituted acyloxy is anacyloxy that is optionally substituted as described herein for acyl.

The term “alcohol oxygen atom,” as used herein, refers to a divalentoxygen atom bonded to an sp³-hybridized carbon atom and to anothersp³-hybridized carbon atom or an sp²-hybridized carbon atom of acarbonyl group.

The term “aldonyl,” as used herein, refers to a monovalent substituentthat is aldonic acid in which a carboxylate hydroxyl is replaced with avalency.

The term “alkanoyl,” as used herein, represents a chemical substituentof formula —C(O)—R, where R is alkyl. An optionally substituted alkanoylis an alkanoyl that is optionally substituted as described herein foralkyl.

The term “alkoxy,” as used herein, represents a chemical substituent offormula —OR, where R is a C₁₋₆ alkyl group, unless otherwise specified.An optionally substituted alkoxy is an alkoxy group that is optionallysubstituted as defined herein for alkyl.

The term “alkenyl,” as used herein, represents acyclic monovalentstraight or branched chain hydrocarbon groups containing one, two, orthree carbon-carbon double bonds. Alkenyl, when unsubstituted, has from2 to 22 carbons, unless otherwise specified. In certain preferredembodiments, alkenyl, when unsubstituted, has from 2 to 12 carbon atoms(e.g., 1 to 8 carbons). Non-limiting examples of the alkenyl groupsinclude ethenyl, prop-1-enyl, prop-2-enyl, 1-methylethenyl, but-1-enyl,but-2-enyl, but-3-enyl, 1-methylprop-1-enyl, 2-methylprop-1-enyl, and1-methylprop-2-enyl. Alkenyl groups may be optionally substituted asdefined herein for alkyl.

The term “alkyl,” as used herein, refers to an acyclic straight orbranched chain saturated hydrocarbon group, which, when unsubstituted,has from 1 to 22 carbons (e.g., 1 to 20 carbons), unless otherwisespecified. In certain preferred embodiments, alkyl, when unsubstituted,has from 1 to 12 carbons (e.g., 1 to 8 carbons). Alkyl groups areexemplified by methyl; ethyl; n- and iso-propyl; n-, sec-, iso- andtert-butyl; neopentyl, and the like, and may be optionally substituted,valency permitting, with one, two, three, or, in the case of alkylgroups of two carbons or more, four or more substituents independentlyselected from the group consisting of: alkoxy; acyloxy; alkylsulfenyl;alkylsulfinyl; alkylsulfonyl; amino; aryl; aryloxy; azido; cycloalkyl;cycloalkoxy; halo; heterocyclyl; heteroaryl; heterocyclylalkyl;heteroarylalkyl; heterocyclyloxy; heteroaryloxy; hydroxy; nitro;thioalkyl; thioalkenyl; thioaryl; thiol; silyl; cyano; ═O; ═S; and ═NR′,where R′ is H, alkyl, aryl, or heterocyclyl. Each of the substituentsmay itself be unsubstituted or, valency permitting, substituted withunsubstituted substituent(s) defined herein for each respective group.

The term “alkenylene,” as used herein, refers to a straight or branchedchain alkenyl group with one hydrogen removed, thereby rendering thisgroup divalent. Non-limiting examples of the alkenylene groups includeethen-1,1-diyl; ethen-1,2-diyl; prop-1-en-1,1-diyl, prop-2-en-1,1-diyl;prop-1-en-1,2-diyl, prop-1-en-1,3-diyl; prop-2-en-1,1-diyl;prop-2-en-1,2-diyl; but-1-en-1,1-diyl; but-1-en-1,2-diyl;but-1-en-1,3-diyl; but-1-en-1,4-diyl; but-2-en-1,1-diyl;but-2-en-1,2-diyl; but-2-en-1,3-diyl; but-2-en-1,4-diyl;but-2-en-2,3-diyl; but-3-en-1,1-diyl; but-3-en-1,2-diyl;but-3-en-1,3-diyl; but-3-en-2,3-diyl; buta-1,2-dien-1,1-diyl;buta-1,2-dien-1,3-diyl; buta-1,2-dien-1,4-diyl; buta-1,3-dien-1,1-diyl;buta-1,3-dien-1,2-diyl; buta-1,3-dien-1,3-diyl; buta-1,3-dien-1,4-diyl;buta-1,3-dien-2,3-diyl; buta-2,3-dien-1,1-diyl; andbuta-2,3-dien-1,2-diyl.

An optionally substituted alkenylene is an alkenylene that is optionallysubstituted as described herein for alkyl.

The term “alkylene,” as used herein, refers to a saturated divalenthydrocarbon group that is a straight or branched chain saturatedhydrocarbon, in which two valencies replace two hydrogen atoms.Non-limiting examples of the alkylene group include methylene,ethane-1,2-diyl, ethane-1,1-diyl, propane-1,3-diyl, propane-1,2-diyl,propane-1,1-diyl, propane-2,2-diyl, butane-1,4-diyl, butane-1,3-diyl,butane-1,2-diyl, butane-1,1-diyl, and butane-2,2-diyl, butane-2,3-diyl.An optionally substituted alkylene is an alkylene that is optionallysubstituted as described herein for alkyl.

The term “alkylsulfenyl,” as used herein, represents a group of formula—S-(alkyl). An optionally substituted alkylsulfenyl is an alkylsulfenylthat is optionally substituted as described herein for alkyl.

The term “alkylsulfinyl,” as used herein, represents a group of formula—S(O)-(alkyl). An optionally substituted alkylsulfinyl is analkylsulfinyl that is optionally substituted as described herein foralkyl.

The term “alkylsulfonyl,” as used herein, represents a group of formula—S(O)₂-(alkyl). An optionally substituted alkylsulfonyl is analkylsulfonyl that is optionally substituted as described herein foralkyl.

The term “amino acid,” as used herein, represents proline, taurine, or acompound having an amino group and a carboxylate or sulfonate groupseparated by an optionally substituted alkylene or optionallysubstituted arylene. Amino acids are small molecules and have amolecular weight of <900 g/mol (preferably, <500 g/mol). Preferably,when the linker is alkylene, the linker may be optionally substituted asdescribed herein for alkyl. In some embodiments, optionally substitutedalkylene is an alkylene substituted with 1 or 2 groups that areindependently hydroxyl, thiol, amino, guanidine, carbamoylamino,imidazolyl, indolyl, —SeH, oxo, 4-hydroxyphenyl, phenyl, or —SMe.Non-limiting examples of amino acids include alanine, arginine,asparagine, aspartic acid, cysteine, glutamic acid, glutamine, glycine,histidine, isoleucine, leucine, lysine, methionine, phenylalanine,proline, selenocysteine, serine, threonine, tyrosine, tryptophan,ornithine, citrulline, aminobenzoic acid, and taurine.

The term “amino acid metabolite,” as used herein, representsproteinogenic amino acids, in which the α-amino group is replaced with—OH or —H, in which the 1-carboxyl group is replaced with H, in whichthe α-(CHNH₂) group is replaced with a carbonyl, in which the α-aminogroup and (3-hydrogen atom are replaced with a double bond, or in whichthe 1-carboxyl group is replaced with hydroxyl and the α-(CHNH₂) groupis replaced with a carbonyl. Non-limiting examples of amino acidmetabolites include indole-3-acetic acid, indole-3-propionic acid,3-(indole-3-yl)-acrylic acid, indole-3-pyruvic acid, and3-(indol-3-yl)-2-hydroxypropionic acid.

The term “amino acid metabolite acyl,” as used herein, represents anamino acid metabolite, in which carboxylate —OH is replaced with avalency.

The term “aryl,” as used herein, represents a mono-, bicyclic, ormulticyclic carbocyclic ring system having one or two aromatic rings.Aryl group may include from 6 to 10 carbon atoms. All atoms within anunsubstituted carbocyclic aryl group are carbon atoms. Non-limitingexamples of carbocyclic aryl groups include phenyl, naphthyl,1,2-dihydronaphthyl, 1,2,3,4-tetrahydronaphthyl, fluorenyl, indanyl,indenyl, etc. The aryl group may be unsubstituted or substituted withone, two, three, four, or five substituents independently selected fromthe group consisting of: alkyl; alkenyl; alkoxy; acyloxy; amino; aryl;aryloxy; azido; cycloalkyl; cycloalkoxy; halo; heterocyclyl; heteroaryl;heterocyclylalkyl; heteroarylalkyl; heterocyclyloxy; heteroaryloxy;hydroxy; nitro; thioalkyl; thioalkenyl; thioaryl; thiol; silyl; andcyano. Each of the substituents may itself be unsubstituted orsubstituted with unsubstituted substituent(s) defined herein for eachrespective group.

The term “aryl alkyl,” as used herein, represents an alkyl groupsubstituted with an aryl group. An optionally substituted aryl alkyl isan aryl alkyl, in which aryl and alkyl portions may be optionallysubstituted as the individual groups as described herein.

The term “aryloxy,” as used herein, represents a group —OR, where R isaryl. Aryloxy may be an optionally substituted aryloxy. An optionallysubstituted aryloxy is aryloxy that is optionally substituted asdescribed herein for aryl.

The term “autoimmune disorder,” as used herein, refers to a group ofdiseases resulting from one's own immune system incorrectly attackingone's own tissue. Non-limiting examples of autoimmune disorders includean inflammatory bowel disease, Addison's disease, alopecia areata,ankylosing spondylitis, antiphospholipid syndrome, hemolytic anemia,autoimmune hepatitis, Behcet's disease, Berger's disease, bullouspemphigoid, cardiomyopathy, celiac sprue, chronic fatigue immunedysfunction syndrome (CFIDS), chronic inflammatory demyelinatingpolyneuropathy, Churg-Strauss syndrome, cicatricial pemphigoid, coldagglutinin disease, type 1 diabetes, discoid lupus, essential mixedcryoglobulinemia, Graves' disease, Guillain-Barré syndrome, Hashimoto'sthyroiditis, hypothyroidism, autoimmune lymphoproliferative syndrome(ALPS), idiopathic pulmonary fibrosis, idiopathic thrombocytopeniapurpura (ITP), juvenile arthritis, lichen planus, lupus erythematosus,Meniere's disease, mixed connective tissue disease, multiple sclerosis,myasthenia gravis, pemphigus vulgaris, pernicious anemia,polychondritis, autoimmune polyglandular syndromes, polymyalgiarheumatica, polymyositis, dermatomyositis, primary agammaglobulinemia,primary biliary cirrhosis, psoriasis, psoriatic arthritis, Raynaud'sphenomenon, Reiter's syndrome, rheumatic fever, rheumatoid arthritis,sarcoidosis, scleroderma, Sjögren's syndrome, stiff-man syndrome,Takayasu arteritis, giant cell arteritis, ulcerative colitis, uveitis,vasculitis, and granulomatosis with polyangiitis. Preferably, theautoimmune disorder is an inflammatory bowel disease (e.g., Crohn'sdisease or ulcerative colitis).

The term “autoimmunity marker,” as used herein, is an observableindication of the presence, absence, or risk of an autoimmune disorder(e.g., an inflammatory bowel disease, Addison's disease, alopeciaareata, ankylosing spondylitis, antiphospholipid syndrome, hemolyticanemia, autoimmune hepatitis, Behcet's disease, Berger's disease,bullous pemphigoid, cardiomyopathy, celiac sprue, chronic fatigue immunedysfunction syndrome (CFIDS), chronic inflammatory demyelinatingpolyneuropathy, Churg-Strauss syndrome, cicatricial pemphigoid, coldagglutinin disease, type 1 diabetes, discoid lupus, essential mixedcryoglobulinemia, Graves' disease, Guillain-Barré syndrome, Hashimoto'sthyroiditis, hypothyroidism, autoimmune lymphoproliferative syndrome(ALPS), idiopathic pulmonary fibrosis, idiopathic thrombocytopeniapurpura (ITP), juvenile arthritis, lichen planus, lupus erythematosus,Meniere's disease, mixed connective tissue disease, multiple sclerosis,myasthenia gravis, pemphigus vulgaris, pernicious anemia,polychondritis, autoimmune polyglandular syndromes, polymyalgiarheumatica, polymyositis, dermatomyositis, primary agammaglobulinemia,primary biliary cirrhosis, psoriasis, psoriatic arthritis, Raynaud'sphenomenon, Reiter's syndrome, rheumatic fever, rheumatoid arthritis,sarcoidosis, scleroderma, Sjögren's syndrome, stiff-man syndrome,Takayasu arteritis, giant cell arteritis, ulcerative colitis, uveitis,vasculitis, and granulomatosis with polyangiitis).

The level of an autoimmune marker may directly or inversely correlatewith an autoimmune disorder state. Non-limiting examples of theautoimmunity markers are a CYP1A1 mRNA level, intestinal motility,CD4⁺CD25⁺ Treg cell (e.g., CD4⁺CD25⁺Foxp3⁺ Treg cell) count, mucussecretion, T_(h)1 cell count, interleukin-8 (IL-8) level, macrophageinflammatory protein 1α (MIP-1α) level, macrophage inflammatory protein1β (MIP-1β) level, NFκB level, inducible nitric oxide synthase (iNOS)level, matrix metallopeptidase 9 (MMP9) level, interferon γ (IFNγ)level, interleukin-17 (IL17) level, intercellular adhesion molecule(ICAM) level, CXCL13 level, 8-iso-prostaglandin F_(2α) (8-iso-PGF2α)level, IgA level, calprotectin level, lipocalin-2 level, short chainfatty acids level, and indoxyl sulfate level.

Autoimmunity markers may be measured using methods known in the art. Forexample, blood sample analyses may be used to measure a CD4⁺CD25⁺ Tregcell (e.g., CD4⁺CD25⁺Foxp3⁺ Treg cell) count, T_(h)1 cell count, NFκBlevel, inducible nitric oxide synthase (iNOS) level, matrixmetallopeptidase 9 (MMP9) level, interferon γ (IFNγ) level,interleukin-17 (IL17) level, intercellular adhesion molecule (ICAM)level, CXCL13 level, and 8-iso-prostaglandin F_(2α) (8-iso-PGF2α) level.Stool sample analyses may be performed to measure an IgA level,calprotectin level, lipocalin-2 level, and short chain fatty acidslevel. Urine sample analysis may be performed to measure an indoxylsulfate level.

The term “carboxylate,” as used herein, represents group —COOH or a saltthereof.

The term “carotenoid,” as used herein, represents a compound of formula:

where

Non-limiting examples of the carotenoid include:

When the carotenoid is acylated, one or both of the hydroxyl groups inthe carotenoid is independently substituted with a group containing afatty acid acyl or a group containing a ketone body or pre-ketone body.

The term “catechin polyphenol,” as used herein, refers to a compound offormula:

where

is a single carbon-carbon bond or double carbon-carbon bond;

Q is —CH₂— or —C(O)—;

each R¹ and each R³ is independently H, halogen, —OR^(A), phosphate, orsulfate;

R² is H or —OR^(A);

each R^(A) is independently H, optionally substituted alkyl, amonosaccharide, a sugar acid, or benzoyl optionally substituted with 1,2, 3, or 4 substituents independently selected from the group consistingof H, hydroxy, halogen, optionally substituted alkyl, optionallysubstituted alkoxy, monosaccharide, sugar acid, phosphate, and sulfate;and

each of n and m is independently 1, 2, 3, or 4.

Non-limiting examples of catechin polyphenols include epigallocatechingallate, quercetin, or myricetin. Preferably, the catechin polyphenol isquercetin. When a catechin polyphenol is acylated, one or more of thehydroxyl groups in the catechin polyphenol are independently substitutedwith a group containing a fatty acid, a group containing a ketone bodyor pre-ketone body, or a group containing an amino acid metabolite.

The expression “C_(x-y),” as used herein, indicates that the group, thename of which immediately follows the expression, when unsubstituted,contains a total of from x to y carbon atoms. If the group is acomposite group (e.g., aryl alkyl), C_(x-y) indicates that the portion,the name of which immediately follows the expression, whenunsubstituted, contains a total of from x to y carbon atoms. Forexample, (C₆₋₁₀-aryl)-C₁₋₆-alkyl is a group, in which the aryl portion,when unsubstituted, contains a total of from 6 to 10 carbon atoms, andthe alkyl portion, when unsubstituted, contains a total of from 1 to 6carbon atoms.

The term “cycloalkyl,” as used herein, refers to a cyclic alkyl grouphaving from three to ten carbons (e.g., a C₃-C₁₀ cycloalkyl), unlessotherwise specified. Cycloalkyl groups may be monocyclic or bicyclic.Bicyclic cycloalkyl groups may be of bicyclo[p.q.0]alkyl type, in whicheach of p and q is, independently, 1, 2, 3, 4, 5, 6, or 7, provided thatthe sum of p and q is 2, 3, 4, 5, 6, 7, or 8. Alternatively, bicycliccycloalkyl groups may include bridged cycloalkyl structures, e.g.,bicyclo[p.q.r]alkyl, in which r is 1, 2, or 3, each of p and q is,independently, 1, 2, 3, 4, 5, or 6, provided that the sum of p, q, and ris 3, 4, 5, 6, 7, or 8. The cycloalkyl group may be a spirocyclic group,e.g., spiro[p.q]alkyl, in which each of p and q is, independently, 2, 3,4, 5, 6, or 7, provided that the sum of p and q is 4, 5, 6, 7, 8, or 9.Non-limiting examples of cycloalkyl include cyclopropyl, cyclobutyl,cyclopentyl, cyclohexyl, cycloheptyl, 1-bicyclo[2.2.1.]heptyl,2-bicyclo[2.2.1.]heptyl, 5-bicyclo[2.2.1]heptyl, 7-bicyclo[2.2.1]heptyl,and decalinyl. The cycloalkyl group may be unsubstituted or substituted(e.g., optionally substituted cycloalkyl) with one, two, three, four, orfive substituents independently selected from the group consisting of:alkyl; alkenyl; alkoxy; acyloxy; alkylsulfenyl; alkylsulfinyl;alkylsulfonyl; amino; aryl; aryloxy; azido; cycloalkyl; cycloalkoxy;halo; heterocyclyl; heteroaryl; heterocyclylalkyl; heteroarylalkyl;heterocyclyloxy; heteroaryloxy; hydroxy; nitro; thioalkyl; thioalkenyl;thioaryl; thiol; silyl; cyano; ═O; ═S; ═NR′, where R′ is H, alkyl, aryl,or heterocyclyl. Each of the substituents may itself be unsubstituted orsubstituted with unsubstituted substituent(s) defined herein for eachrespective group.

The term “cycloalkylene,” as used herein, represents a divalent groupthat is a cycloalkyl group, in which one hydrogen atom is replaced witha valency. An optionally substituted cycloalkylene is a cycloalkylenethat is optionally substituted as described herein for cycloalkyl.

The term “cycloalkoxy,” as used herein, represents a group —OR, where Ris cycloalkyl. An optionally substituted cycloalkoxy is cycloalkoxy thatis optionally substituted as described herein for cycloalkyl.

The term “dialkylamino,” as used herein, refers to a group —NR₂, whereeach R is independently alkyl.

The terms “ellagic acid” and “ellagic acid analogue,” as used herein,collectively refer to a compound of the structure:

where

each of R², R³, and R⁴ is independently H or —OR^(A);

R⁶ is H or —(CO)—R^(5B);

R^(1A) is H or —OR^(A), and R^(5A) is —OH or —OR^(A); or R^(1A) andR^(5A) combine to form —O—;

R^(1B) is H or —OR^(A), and R^(5B) is absent, —OH, or —OR^(A); or R^(1B)and R^(5B) combine to form —O—;

each R^(A) is independently H or O-protecting group.

When the ellagic acid or its analogue is present in an acylated ellagicacid or an acylated ellagic acid analogue, from one to all hydroxyls inthe ellagic acid or its analogue are substituted with a group containinga fatty acid, a group containing a ketone body or pre-ketone body, or agroup containing an amino acid metabolite. The term “ellagic acidanalogue,” refers to the compounds and groups of the above structurethat are not ellagic acid. The term “ellagic acid” refers to thefollowing two compounds:

or these compounds within the structure of an acylated ellagic acid.

Non-limiting examples of ellagic acid analogues include urolithin A,urolithin B, urolithin C, urolithin D, urolithin E, and urolithin M5.

The term “ester bond,” as used herein, refers to a covalent bond betweenan alcohol or phenolic oxygen atom and a carbonyl group that is furtherbonded to a carbon atom.

The term “fatty acid,” as used herein, refers to a short-chain fattyacid, a medium chain fatty acid, a long chain fatty acid, a very longchain fatty acid, or an unsaturated analogue thereof, or aphenyl-substituted analogue thereof. Short chain fatty acids containfrom 1 to 6 carbon atoms, medium chain fatty acids contain from 7 to 13carbon atoms, and a long-chain fatty acids contain from 14 to 22 carbonatoms. A fatty acid may be saturated or unsaturated. An unsaturatedfatty acid includes 1, 2, 3, 4, 5, or 6 carbon-carbon double bonds.Preferably, the carbon-carbon double bonds in unsaturated fatty acidshave Z stereochemistry.

The term “fatty acid acyl,” as used herein, refers to a fatty acid, inwhich the hydroxyl group is replaced with a valency.

The term “fatty acid acyloxy,” as used herein, refers to group —OR,where R is a fatty acid acyl.

The term “a group containing an amino acid metabolite,” as used herein,represents a monovalent substituent including at least one amino acidmetabolite within its structure and having the valency on a carbon atomof a carbonyl group or on an anomeric carbon atom. A group containing anamino acid metabolite bonds to a core through a carbonate linker,carbamate linker, ester bond, glycosidic bond, or amide bond. A groupcontaining an amino acid metabolite may be a group selected from thegroup consisting of monosaccharide, ketone body, pre-ketone body,aldonyl, uronyl, ulosonyl, and amino acid metabolite acyl, and whereeach hydroxyl in the monosaccharide, ketone body, pre-ketone body,aldonyl, uronyl, and ulosonyl is optionally and independentlysubstituted with an amino acid metabolite acyl.

The term “group containing a fatty acid,” as used herein, represents amonovalent substituent including at least one fatty acid within itsstructure and having the valency on a carbon atom of a carbonyl group oron an anomeric carbon atom. A group containing a fatty acid bonds to acore through a carbonate linker, carbamate linker, ester bond,glycosidic bond, or amide bond. A group containing a fatty acid may be agroup selected from the group consisting of monosaccharide, ketone body,pre-ketone body, aldonyl, uronyl, ulosonyl, and fatty acid acyl, andwhere each hydroxyl in the monosaccharide, ketone body, pre-ketone body,aldonyl, uronyl, and ulosonyl is optionally and independentlysubstituted with a fatty acid acyl.

The term “group containing a ketone body or pre-ketone body,” as usedherein, represents a monovalent substituent including at least oneketone body and/or at least one pre-ketone body within its structure andhaving the valency on a carbon atom of a carbonyl group or on ananomeric carbon atom. A group containing a ketone body or pre-ketonebody bonds to a core through a carbonate linker, ester bond, orglycosidic bond. A group containing a ketone body or pre-ketone body maybe a group selected from the group consisting of monosaccharide, ketonebody, aldonyl, uronyl, ulosonyl, and —C(O)—R, where R is a pre-ketonebody or ketone body, and where each hydroxyl in the monosaccharide,ketone body, pre-ketone body, aldonyl, uronyl, and ulosonyl isoptionally and independently substituted with an acyl or ketone body, ahydroxyl group in which, if present, is optionally substituted with anacyl.

The term “halogen,” as used herein, represents a halogen selected frombromine, chlorine, iodine, and fluorine.

The term “heteroaryl,” as used herein, represents a monocyclic 5-, 6-,7-, or 8-membered ring system, or a fused or bridging bicyclic,tricyclic, or tetracyclic ring system; the ring system contains one,two, three, or four heteroatoms independently selected from the groupconsisting of nitrogen, oxygen, and sulfur; and at least one of therings is an aromatic ring. Non-limiting examples of heteroaryl groupsinclude benzimidazolyl, benzofuryl, benzothiazolyl, benzothienyl,benzoxazolyl, furyl, imidazolyl, indolyl, isoindazolyl, isoquinolinyl,isothiazolyl, isothiazolyl, isoxazolyl, oxadiazolyl, oxazolyl, purinyl,pyrrolyl, pyridinyl, pyrazinyl, pyrimidinyl, qunazolinyl, quinolinyl,thiadiazolyl (e.g., 1,3,4-thiadiazole), thiazolyl, thienyl, triazolyl,tetrazolyl, dihydroindolyl, tetrahydroquinolyl, tetrahydroisoquinolyl,etc. The term bicyclic, tricyclic, and tetracyclic heteroaryls includeat least one ring having at least one heteroatom as described above andat least one aromatic ring. For example, a ring having at least oneheteroatom may be fused to one, two, or three carbocyclic rings, e.g.,an aryl ring, a cyclohexane ring, a cyclohexene ring, a cyclopentanering, a cyclopentene ring, or another monocyclic heterocyclic ring.Examples of fused heteroaryls include 1,2,3,5,8,8a-hexahydroindolizine;2,3-dihydrobenzofuran; 2,3-dihydroindole; and 2,3-dihydrobenzothiophene.Heteroaryl may be optionally substituted with one, two, three, four, orfive substituents independently selected from the group consisting of:alkyl; alkenyl; alkoxy; acyloxy; aryloxy; alkylsulfenyl; alkylsulfinyl;alkylsulfonyl; amino; arylalkoxy; cycloalkyl; cycloalkoxy; halogen;heterocyclyl; heterocyclyl alkyl; heteroaryl; heteroaryl alkyl;heterocyclyloxy; heteroaryloxy; hydroxyl; nitro; thioalkyl; thioalkenyl;thioaryl; thiol; cyano; ═O; —NR2, where each R is independentlyhydrogen, alkyl, acyl, aryl, arylalkyl, cycloalkyl, heterocyclyl, orheteroaryl; —COOR^(A), where R^(A) is hydrogen, alkyl, aryl, arylalkyl,cycloalkyl, heterocyclyl, or heteroaryl; and —CON(R^(B))₂, where eachR^(B) is independently hydrogen, alkyl, aryl, arylalkyl, cycloalkyl,heterocyclyl, or heteroaryl. Each of the substituents may itself beunsubstituted or substituted with unsubstituted substituent(s) definedherein for each respective group.

The term “heteroaryloxy,” as used herein, refers to a structure —OR, inwhich R is heteroaryl. Heteroaryloxy can be optionally substituted asdefined for heteroaryl.

The term “heterocyclyl,” as used herein, represents a monocyclic,bicyclic, tricyclic, or tetracyclic non-aromatic ring system havingfused or bridging 4-, 5-, 6-, 7-, or 8-membered rings, unless otherwisespecified, the ring system containing one, two, three, or fourheteroatoms independently selected from the group consisting ofnitrogen, oxygen, and sulfur. Non-aromatic 5-membered heterocyclyl haszero or one double bonds, non-aromatic 6- and 7-membered heterocyclylgroups have zero to two double bonds, and non-aromatic 8-memberedheterocyclyl groups have zero to two double bonds and/or zero or onecarbon-carbon triple bond. Heterocyclyl groups have a carbon count of 1to 16 carbon atoms unless otherwise specified. Certain heterocyclylgroups may have a carbon count up to 9 carbon atoms. Non-aromaticheterocyclyl groups include pyrrolinyl, pyrrolidinyl, pyrazolinyl,pyrazolidinyl, imidazolinyl, imidazolidinyl, piperidinyl,homopiperidinyl, piperazinyl, pyridazinyl, oxazolidinyl,isoxazolidiniyl, morpholinyl, thiomorpholinyl, thiazolidinyl,isothiazolidinyl, thiazolidinyl, tetrahydrofuranyl, dihydrofuranyl,tetrahydrothienyl, dihydrothienyl, pyranyl, dihydropyranyl, dithiazolyl,etc. The term “heterocyclyl” also represents a heterocyclic compoundhaving a bridged multicyclic structure in which one or more carbonsand/or heteroatoms bridges two non-adjacent members of a monocyclicring, e.g., quinuclidine, tropanes, or diaza-bicyclo[2.2.2]octane. Theterm “heterocyclyl” includes bicyclic, tricyclic, and tetracyclic groupsin which any of the above heterocyclic rings is fused to one, two, orthree carbocyclic rings, e.g., a cyclohexane ring, a cyclohexene ring, acyclopentane ring, a cyclopentene ring, or another heterocyclic ring.Examples of fused heterocyclyls include1,2,3,5,8,8a-hexahydroindolizine; 2,3-dihydrobenzofuran;2,3-dihydroindole; and 2,3-dihydrobenzothiophene. The heterocyclyl groupmay be unsubstituted or substituted with one, two, three, four or fivesubstituents independently selected from the group consisting of: alkyl;alkenyl; alkoxy; acyloxy; alkylsulfenyl; alkylsulfinyl; alkylsulfonyl;aryloxy; amino; arylalkoxy; cycloalkyl; cycloalkoxy; halogen;heterocyclyl; heterocyclyl alkyl; heteroaryl; heteroaryl alkyl;heterocyclyloxy; heteroaryloxy; hydroxyl; nitro; thioalkyl; thioalkenyl;thioaryl; thiol; cyano; ═O; ═S; —NR₂, where each R is independentlyhydrogen, alkyl, acyl, aryl, arylalkyl, cycloalkyl, heterocyclyl, orheteroaryl; —COOR^(A), where R^(A) is hydrogen, alkyl, aryl, arylalkyl,cycloalkyl, heterocyclyl, or heteroaryl; and —CON(R^(B))₂, where eachR^(B) is independently hydrogen, alkyl, aryl, arylalkyl, cycloalkyl,heterocyclyl, or heteroaryl.

The term “heterocyclyl alkyl,” as used herein, represents an alkyl groupsubstituted with a heterocyclyl group. The heterocyclyl and alkylportions of an optionally substituted heterocyclyl alkyl are optionallysubstituted as the described for heterocyclyl and alkyl, respectively.

The term “heterocyclylene,” as used herein, represents a heterocyclyl,in which one hydrogen atom is replaced with a valency. An optionallysubstituted heterocyclylene is a heterocyclylene that is optionallysubstituted as described herein for heterocyclyl.

The term “heterocyclyloxy,” as used herein, refers to a structure —OR,in which R is heterocyclyl. Heterocyclyloxy can be optionallysubstituted as described for heterocyclyl.

The term “hydroxybenzoic acid,” as used herein, represents a compound ofthe following structure:

or a salt thereof,

where

n is 1, 2, or 3;

each R¹ is independently H or alkyl; and

R² is H or alkyl.

Non-limiting examples of hydroxybenzoic acids include salicylic acid andgallic acid.

The terms “hydroxyl” and “hydroxy,” as used interchangeably herein,represent —OH.

The term “ketone body,” as used herein, refers to (i) p-hydroxybutyricacid, or (ii) a group that is β-hydroxy butyric acid, where at least onehydroxyl hydrogen atom is replaced with a valency or a carboxylate —OHis replaced with a valency.

The term “ketone body acyl,” as used herein, refers to ap-hydroxybutyric acid, in which the carboxylate —OH group is replacedwith a valency.

The term “4-methyl-1,3-dioxan-2-yl,” as used herein, refers to themonovalent group of formula:

where R¹ is optionally substituted C₁₋₆ alkyl (e.g., methyl).

The term “modulating,” as used herein, refers to an observable change inthe level of a marker in a subject, as measured using techniques andmethods known in the art for the measurement of the marker. Modulatingthe marker level in a subject may result in a change of at least 1%relative to prior to administration (e.g., at least 5%, 10%, 15%, 20%,25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%,95% or at least 98% or more relative to prior to administration; e.g.,up to 100% relative to prior to administration). In some embodiments,modulating is increasing the level of a marker in a subject. Increasingthe marker level in a subject may result in an increase of at least 1%relative to prior to administration (e.g., at least 5%, 10%, 15%, 20%,25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%,95% or at least 98% or more relative to prior to administration; e.g.,up to 100% relative to prior to administration). In other embodiments,modulating is decreasing the level of a marker in a subject. Decreasingthe marker level in a subject may result in a decrease of at least 1%relative to prior to administration (e.g., at least 5%, 10%, 15%, 20%,25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%,95% or at least 98% or more relative to prior to administration; e.g.,up to 100% relative to prior to administration). In embodiments in whicha parameter is increased or decreased (or reduced) in a subjectfollowing a step of administering a composition described herein, theincrease or decrease may take place and/or be detectable within a rangeof time following the administration (e.g., within six hours, 24 hours,3 days, a week or longer), and may take place and/or be detectable afterone or more administrations (e.g., after 2, 3, 4, 5, 6, 7, 8, 9, 10 ormore administrations, e.g., as part of a dosing regimen for thesubject).

The term “oxo,” as used herein, represents a divalent oxygen atom (e.g.,the structure of oxo may be shown as ═O).

The term “phenolic oxygen atom,” as used herein, refers to a divalentoxygen atom within the structure of a compound, where one valency of thephenolic oxygen atom is bonded to a first carbon atom, and anothervalency is bonded to a second carbon atom, where the first carbon atomis an sp²-hybridized carbon atom within a benzene ring, and the secondcarbon atom is an sp³-hybridized carbon atom or an sp²-hybridized carbonatom.

The term “phosphate, as used herein, represents group —OPO(OH)₂ or asalt thereof.

The term “pre-ketone body,” as used herein, represents (i) a ketone bodyhaving hydroxymethyl instead of a carboxylate, or (ii) a group that is aketone body having hydroxymethyl instead of a carboxylate, where atleast one hydroxyl is replaced with —OR, where R is a valency. Anon-limiting example of a pre-ketone body is butane-1,3-diol or4-hydroxybutan-2-one. The term “pre-ketone body,” as used herein, alsorepresents (4-methyl-1,3-dioxan-2-yl)-(alkylene)_(n)-CO—R^(A), where nis 0 or 1, and R^(A) is —OH, if the pre-ketone body is not part of anacylated active agent, or a valency if the pre-ketone body is part of agroup including a pre-ketone body (e.g., a pre-ketone body acyl). Anon-limiting example of a pre-ketone body is butane-1,3-diol or4-hydroxybutan-2-one

The term “pre-ketone body acyl,” as used herein, refers to a pre-ketonebody, in which the carboxylate —OH group is replaced with a valency.

The term “protecting group,” as used herein, represents a group intendedto protect a hydroxy, an amino, or a carbonyl from participating in oneor more undesirable reactions during chemical synthesis. The term“O-protecting group,” as used herein, represents a group intended toprotect a hydroxy or carbonyl group from participating in one or moreundesirable reactions during chemical synthesis. The term “N-protectinggroup,” as used herein, represents a group intended to protect anitrogen containing (e.g., an amino or hydrazine) group fromparticipating in one or more undesirable reactions during chemicalsynthesis. Commonly used O- and N-protecting groups are disclosed inGreene, “Protective Groups in Organic Synthesis,” 3^(rd) Edition (JohnWiley & Sons, New York, 1999), which is incorporated herein byreference. Exemplary O- and N-protecting groups include alkanoyl,aryloyl, or carbamyl groups such as formyl, acetyl, propionyl, pivaloyl,t-butylacetyl, 2-chloroacetyl, 2-bromoacetyl, trifluoroacetyl,trichloroacetyl, phthalyl, o-nitrophenoxyacetyl, α-chlorobutyryl,benzoyl, 4-chlorobenzoyl, 4-bromobenzoyl, f-butyldimethylsilyl,tri-/so-propylsilyloxymethyl, 4,4′-dimethoxytrityl, isobutyryl,phenoxyacetyl, 4-isopropylpehenoxyacetyl, dimethylformamidino, and4-nitrobenzoyl.

Exemplary O-protecting groups for protecting carbonyl containing groupsinclude, but are not limited to: acetals, acylals, 1,3-dithianes,1,3-dioxanes, 1,3-dioxolanes, and 1,3-dithiolanes.

Other O-protecting groups include, but are not limited to: substitutedalkyl, aryl, and aryl-alkyl ethers (e.g., trityl; methylthiomethyl;methoxymethyl; benzyloxymethyl; siloxymethyl;2,2,2,-trichloroethoxymethyl; tetrahydropyranyl; tetrahydrofuranyl;ethoxyethyl; 1-[2-(trimethylsilyl)ethoxy]ethyl; 2-trimethylsilylethyl;t-butyl ether; p-chlorophenyl, p-methoxyphenyl, p-nitrophenyl, benzyl,p-methoxybenzyl, and nitrobenzyl); silyl ethers (e.g., trimethylsilyl;triethylsilyl; triisopropylsilyl; dimethylisopropylsilyl;t-butyldimethylsilyl; t-butyldiphenylsilyl; tribenzylsilyl;triphenylsilyl; and diphenymethylsilyl); carbonates (e.g., methyl,methoxymethyl, 9-fluorenylmethyl; ethyl; 2,2,2-trichloroethyl;2-(trimethylsilyl)ethyl; vinyl, allyl, nitrophenyl; benzyl;methoxybenzyl; 3,4-dimethoxybenzyl; and nitrobenzyl).

Other N-protecting groups include, but are not limited to, chiralauxiliaries such as protected or unprotected D, L or D, L-amino acidssuch as alanine, leucine, phenylalanine, and the like;sulfonyl-containing groups such as benzenesulfonyl, p-toluenesulfonyl,and the like; carbamate forming groups such as benzyloxycarbonyl,p-chlorobenzyloxycarbonyl, p-methoxybenzyloxycarbonyl,p-nitrobenzyloxycarbonyl, 2-nitrobenzyloxycarbonyl,p-bromobenzyloxycarbonyl, 3,4-dimethoxybenzyloxycarbonyl,3,5-dimethoxybenzyl oxycarbonyl, 2,4-dimethoxybenzyloxycarbonyl,4-methoxybenzyloxycarbonyl, 2-nitro-4,5-dimethoxybenzyloxycarbonyl,3,4,5-trimethoxybenzyloxycarbonyl,1-(p-biphenylyl)-1-methylethoxycarbonyl,α,α-dimethyl-3,5-dimethoxybenzyloxycarbonyl, benzhydryloxy carbonyl,t-butyloxycarbonyl, diisopropylmethoxycarbonyl, isopropyloxycarbonyl,ethoxycarbonyl, methoxycarbonyl, allyloxycarbonyl,2,2,2,-trichloroethoxycarbonyl, phenoxycarbonyl, 4-nitrophenoxycarbonyl, fluorenyl-9-methoxycarbonyl, cyclopentyloxycarbonyl,adamantyloxycarbonyl, cyclohexyloxycarbonyl, phenylthiocarbonyl, and thelike, aryl-alkyl groups such as benzyl, triphenylmethyl,benzyloxymethyl, and the like and silyl groups such as trimethylsilyl,and the like. Useful N-protecting groups are formyl, acetyl, benzoyl,pivaloyl, t-butylacetyl, alanyl, phenylsulfonyl, benzyl,t-butyloxycarbonyl (Boc), and benzyloxycarbonyl (Cbz).

The term “subject,” as used herein, represents a human or non-humananimal (e.g., a mammal) that is suffering from, or is at risk of,disease, disorder, or condition, as determined by a qualifiedprofessional (e.g., a doctor or a nurse practitioner) with or withoutknown in the art laboratory test(s) of sample(s) from the subject.Non-limiting examples of diseases, disorders, and conditions includeautoimmune disorders, as described herein.

The term “sugar acid,” as used herein, refers to a monosaccharide, inthe linear form of which, one or both terminal positions are oxidized toa carboxylic acid. There are four classes of sugar acids: aldonic acid,ulosonic acid, uronic acid, and aldaric acid. Any of the four sugar acidclasses may be used in acylated active agent disclosed herein.Non-limiting examples of sugar acids include gluconic acid.

The term “sulfate,” as used herein, represents group —OSO3H or a saltthereof.

The term “thioalkenyl,” as used herein, represents a group —SR, where Ris alkenyl. An optionally substituted thioalkenyl is thioalkenyl that isoptionally substituted as described herein for alkenyl.

The term “thioalkyl,” as used herein, represents a group —SR, where R isalkyl. An optionally substituted thioalkyl is thioalkyl that isoptionally substituted as described herein for alkyl.

The term “thioaryl,” as used herein, represents a group —SR, where R isaryl. An optionally substituted thioaryl is thioaryl that is optionallysubstituted as described herein for aryl.

“Treatment” and “treating,” as used herein, refer to the medicalmanagement of a subject with the intent to improve, ameliorate,stabilize, prevent or cure a disease, disorder, or condition. This termincludes active treatment (treatment directed to improve the disease,disorder, or condition); causal treatment (treatment directed to thecause of the associated disease, disorder, or condition); palliativetreatment (treatment designed for the relief of symptoms of the disease,disorder, or condition); preventative treatment (treatment directed tominimizing or partially or completely inhibiting the development of theassociated disease, disorder, or condition); and supportive treatment(treatment employed to supplement another therapy).

The term “ulosonyl,” as used herein, refers to a monovalent substituentthat is a ulosonic acid in which a carboxylate hydroxyl is replaced witha valency.

The term “uronyl,” as used herein, refers to a monovalent substituentthat is a uronic acid in which a carboxylate hydroxyl is replaced with avalency.

The compounds described herein, unless otherwise noted, encompassisotopically enriched compounds (e.g., deuterated compounds), tautomers,and all stereoisomers and conformers (e.g. enantiomers, diastereomers,E/Z isomers, atropisomers, etc.), as well as racemates thereof andmixtures of different proportions of enantiomers or diastereomers, ormixtures of any of the foregoing forms as well as salts (e.g.,pharmaceutically acceptable salts).

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is a chart showing the CD4⁺CD25⁺ cell counts as a percentage ofall CD4⁺ T cells in seven animal cohorts: (1) untreated animalsreceiving a normal diet, (2) untreated animals receiving a high-fatdiet, (3) animals receiving acetate along with a high-fat diet, (4)animals receiving epigallocatechin gallate (EGCG) along with a high-fatdiet, (5) animals receiving epigallocatechin gallate (EGCG) and acetateas different compounds along with a high-fat diet, (6) animals receivingepigallocatechin gallate octaacetate (EGCG-8A) along with a high-fatdiet, and (7) animals receiving rosiglitazone along with a high-fatdiet.

FIG. 2 is a chart showing changes in the mucosal thickness in micefollowing the treatment with a vehicle control (methylcellulose),cyclosporine A, or compound 44.

FIG. 3A is a chart showing the colon lengths measured in colon samplesfrom adoptive T-cell transfer mice treated with a vehicle(methyl-cellulose), anti-IL12 antibody (positive control), olsalazine(positive treatment group), or compound 44 (test treatment group). Dataare presented as mean±SEM; *p<0.05 and **p<0.01 were considered to bestatistically significant.

FIG. 3B is a chart showing the colon weight per unit length measured incolon samples from mice treated with a vehicle (methyl-cellulose),anti-IL12 antibody (positive control), olsalazine (positive treatmentgroup), or compound 44 (test treatment group). Data are presented asmean±SEM; *p<0.05 and ***p<0.001 were considered to be statisticallysignificant.

FIG. 4 is a graph showing oral exposure of mesalamine and d5-butyrate inthe colon of mice administered compound 44.

DETAILED DESCRIPTION

The invention provides acylated active agents (e.g., acylated catechinpolyphenols, acylated carotenoids, acylated mesalamines, acylatedsugars, acylated hydroxybenzoic acids, acylated ellagic acid, acylatedellagic acid analogues, and acylated shikimic acids) and methods formodulating an autoimmunity marker in a subject. Without wishing to bebound by theory, the disclosed acylated active agents are believed toact in concert with, or in lieu of, the microbiota of a subject tomodulate, for example, the host's immune system.

It has been surprisingly found that administration of an acylated activeagent (e.g., acylated catechin polyphenol (e.g.,epigallocatechin-3-gallate octaacetate)) to a subject can induce Tregdifferentiation (e.g., CD4⁺CD25⁺ Treg differentiation) and thus canproduce beneficial effects in subjects having an autoimmune disorder.

The components of the acylated active agent (e.g., acylated catechinpolyphenol (e.g., short chain fatty acid acyls (e.g., acetyl) andepigallocatechin gallate)) may act synergistically to modulate anautoimmunity marker, e.g., upon hydrolysis in the GI tract of thesubject receiving the acylated catechin polyphenol. The components ofthe acylated catechin polyphenol (e.g., short chain fatty acid acyls(e.g., acetyl) and epigallocatechin gallate) may act synergistically totreat an autoimmune disorder, e.g., upon hydrolysis in the GI tract ofthe subject receiving the acylated catechin polyphenol.

Advantageously, acylated active agents (e.g., acylated catechinpolyphenols) disclosed herein may have superior organoleptic properties(e.g., palatability). This provides an important advantage as theindividual components (e.g., acetic acid or epigallocatechin gallate)may exhibit less desirable organoleptic properties (e.g., palatability).Improved organoleptic properties facilitate oral administration, and areparticularly advantageous for delivery of high unit dosages (e.g., unitdosages of 0.5 g or higher).

Acylated Active Agents

An acylated active agent disclosed herein may be an acylated catechinpolyphenol, acylated carotenoid, acylated mesalamine, acylated shikimicacid, acylated sugar, acylated ellagic acid, acylated ellagic acidanalogue, or acylated hydroxybenzoic acid.

Typically, an acylated active agent includes a core of formula (A)(e.g., a catechin polyphenol core) linked to at least one group (e.g., agroup containing a fatty acid, a group containing a ketone body orpre-ketone body, or a group containing an amino acid metabolite) throughester bond(s), amide bond(s), carbonate linker(s), carbamate linker(s),and/or glycosidic bond(s). For example, an acylated active agent mayinclude a catechin polyphenol substituted with one or more substituentsindependently selected from the group consisting of an alkyl, acyl,group containing a fatty acid (e.g., a short chain fatty acid or amedium chain fatty acid), group containing a ketone body or pre-ketonebody, and group containing an amino acid metabolite. The fatty acid maybe, e.g., a short chain fatty acid (e.g., acetyl, propionyl, orbutyryl). In some embodiments, the short chain fatty acid is acetyl. Inparticular embodiments, the short chain fatty acid is butyryl.

An acylated active agent disclosed herein may include, e.g., at leastone group containing a fatty acid, group containing a ketone body orpre-ketone body, or group containing an amino acid metabolite.

A group containing a fatty acid may be, e.g., a fatty acid acyl (e.g.,short chain fatty acid or medium chain fatty acid), a monosaccharidehaving one or more hydroxyl groups substituted with fatty acid acyls(e.g., short chain fatty acid acyls or medium chain fatty acid acyls),or a sugar acid (e.g., aldonic acid) having one or more alcohol hydroxylgroups substituted with fatty acid acyls (e.g., short chain fatty acidacyls or medium chain fatty acid acyls). A monosaccharides may be, e.g.,arabinose, xylose, fructose, galactose, glucose, glucosinolate, ribose,tagatose, fucose, or rhamnose. In some embodiments, the monosaccharideis L-arabinose, D-xylose, fructose, galactose, D-glucose, glucosinolate,D-ribose, D-tagatose, L-fucose, or L-rhamnose (e.g., the monosaccharideis D-xylose). The group containing a fatty acid may be, e.g., a fattyacid acyl.

A group containing an amino acid metabolite may be, e.g., an amino acidmetabolite acyl (e.g., indole-3-acetyl, indole-3-acryloyl, orindole-3-pyruvyl), a monosaccharide having one or more hydroxyl groupssubstituted with amino acid metabolite acyls (e.g., indole-3-acetyl,indole-3-acryloyl, or indole-3-pyruvyl), or a sugar acid (e.g., aldonicacid) having one or more alcohol hydroxyl groups substituted with aminoacid metabolite acyls (e.g., indole-3-acetyl, indole-3-acryloyl, orindole-3-pyruvyl). A monosaccharide may be, e.g., arabinose, xylose,fructose, galactose, glucose, glucosinolate, ribose, tagatose, fucose,or rhamnose. In some embodiments, the monosaccharide is L-arabinose,D-xylose, fructose, galactose, D-glucose, glucosinolate, D-ribose,D-tagatose, L-fucose, or L-rhamnose (e.g., the monosaccharide isD-xylose). The group containing an amino acid metabolite may be, e.g.,an amino acid metabolite acyl.

A group containing a ketone body or pre-ketone body may be, e.g., aketone body acyl (e.g., p-hydroxybutyrate acyl), a pre-ketone body acyl,a monosaccharide having one or more hydroxyl groups substituted withketone body acyls (e.g., p-hydroxybutyrate acyl) and/or pre-ketone bodyacyls, or a sugar acid (e.g., aldonic acid) having one or more alcoholhydroxyl groups substituted with ketone body acyls (e.g.,p-hydroxybutyrate acyl) and/or pre-ketone body acyls. A monosaccharidemay be, e.g., arabinose, xylose, fructose, galactose, glucose,glucosinolate, ribose, tagatose, fucose, or rhamnose. In someembodiments, the monosaccharide is L-arabinose, D-xylose, fructose,galactose, D-glucose, glucosinolate, D-ribose, D-tagatose, L-fucose, orL-rhamnose (e.g., the monosaccharide is D-xylose).

The group containing a ketone body or pre-ketone body may be, e.g., aketone body acyl or a pre-ketone body acyl. Preferably, the groupcontaining a ketone body or pre-ketone body is a group containing aketone body (e.g., a group containing p-hydroxybutyrate).

In certain embodiments, the group may be a monovalent group of thefollowing formula:

where

L is absent, carbamate linker, or carbonate linker;

group A is a fatty acid acyl, ketone body, pre-ketone body, amino acidmetabolite acyl, monosaccharide, or sugar acid;

each R is independently ketone body optionally having a hydroxyl groupthat is optionally substituted with an acyl (e.g., a fatty acid acyl),pre-ketone body optionally having a hydroxyl group that is optionallysubstituted with an acyl (e.g., a fatty acid acyl), or acyl (e.g., afatty acid acyl, a ketone body acyl, a pre-ketone body acyl, or an aminoacid metabolite acyl); and

m is an integer from 0 to the total number of available hydroxyl groupsin group A (e.g., 1, 2, 3, 4, or 5);

provided that

L is a carbonate linker or carbamate linker, if group A has a valency ona non-glycosidic alcohol oxygen atom; and

L is absent, if group A has a valency on a carbonyl carbon atom.

In certain embodiments, the group of formula (B) includes at least onefatty acid acyl and/or at least one ketone body and/or at least onepre-ketone body and/or at least one amino acid metabolite.

In some embodiments, the fatty acid(s) are short chain fatty acid acyls(e.g., butyryls). In particular embodiments, the fatty acid(s) in thegroup containing a fatty acid are medium chain fatty acid acyls (e.g.,octanoyl).

Non-limiting examples of the groups are:

where

R is H, —CH₃, or —CH₂OR^(FA); and

each R^(FA) is independently H, a fatty acid acyl (e.g., a short chainfatty acid acyl or medium chain fatty acid acyl), a ketone body acyl(e.g., (3-hydroxybutyrate acyl), a pre-ketone body acyl, or an aminoacid metabolite acyl (e.g., indole-3-acetyl, indole-3-acyloyl, orindole-3-pyruvyl).

Acylated Catechin Polyphenols

An acylated catechin polyphenol of the invention may be a substitutedcompound having the core of formula (A):

or a multimer thereof, or a salt thereof,

where the substituents are independently selected from the groupconsisting of —OR^(A), —OCOO—R^(A), —NHR^(B), oxo, halogen, optionallysubstituted C₁₋₂₀ alkyl, optionally substituted C₂₋₂₀ alkenyl,optionally substituted thioalkyl, optionally substituted alkylsulfonyl,optionally substituted alkylsulfenyl, optionally substitutedalkylsulfinyl, optionally substituted thioaryl, optionally substitutedaryl thioalkyl, optionally substituted thioalkenyl, dialkylamino,sulfate, phosphate, ascorbic acid, optionally substituted heterocyclyl,nitro, amino acids, C₁₋₆ esters of amino acids, optionally acylatedmonosaccharide, and optionally acylated sugar acid, where each R^(A) isindependently H, optionally substituted alkyl, a group containing afatty acid, a group containing a ketone body or pre-ketone body, a groupcontaining an amino acid metabolite, or benzoyl optionally substitutedwith one, two, three, or four substituents independently selected fromthe group consisting of H, hydroxyl, halogen, a group containing a fattyacid, a group containing a ketone body or pre-ketone body, a groupcontaining an amino acid metabolite, optionally substituted alkoxy, andoptionally substituted alkyl, and where R^(B) is independently H oroptionally substituted alkyl;

where the carbon-carbon bond connecting carbon 2 and carbon 3 in formula(A) is a single bond or a double bond;

where the multimer includes a total of 2 or 3 cores of formula (A), eachcore substituted independently as described above; and

where two vicinal centers in core (A) may be further substituted with agroup —(O)_(q)-L¹-L²-, where q is 0 or 1, L¹ is optionally substitutedalkylene, optionally substituted alkenylene, or optionally substitutedheterocyclylene; and L² is a covalent bond, optionally substitutedheterocyclylene, or optionally substituted cycloalkylene.

In some embodiments, at least one of positions 5, 6, 7, and 8 is—OR^(A), where R^(A) is a group containing a fatty acid, groupcontaining a ketone body or pre-ketone body, group containing an aminoacid metabolite, or benzoyl optionally substituted with one, two, three,or four substituents independently selected from the group consisting ofH, hydroxyl, halogen, a group containing a fatty acid, group containinga ketone body or pre-ketone body, group containing an amino acidmetabolite, optionally substituted alkoxy, and optionally substitutedalkyl. In some embodiments, the compound of formula (A) includes atleast one group containing a fatty acid. In some embodiments, thecompound of formula (A) includes at least one group containing a ketonebody or pre-ketone body. In some embodiments, the compound of formula(A) includes at least one group containing an amino acid metabolite.

An acylated catechin polyphenol of the invention may be a catechinpolyphenol, in which one or more hydroxyl groups are independentlyreplaced with —OR, where each R is independently selected from the groupconsisting of an acyl, alkyl, group containing a fatty acid, groupcontaining a ketone body or pre-ketone body, and group containing anamino acid metabolite. In some embodiments, at least one R is a groupcontaining a fatty acid. In some embodiments, at least one R is a groupcontaining a ketone body or pre-ketone body. In some embodiments, atleast one R is a group containing an amino acid metabolite.

An acylated catechin polyphenol may be a compound of formula (I):

or a pharmaceutically acceptable salt thereof, where

is a single carbon-carbon bond or double carbon-carbon bond;

Q is —CH₂— or —C(O)—;

each R¹ and each R³ is independently H, halogen, —OR^(A), phosphate, orsulfate;

R² is H or —OR^(A);

each R^(A) is independently H, optionally substituted alkyl, amonosaccharide, a monosaccharide, a sugar acid, a group containing afatty acid, a group containing a ketone body or pre-ketone body, a groupcontaining an amino acid metabolite, or benzoyl optionally substitutedwith 1, 2, 3, or 4 substituents independently selected from the groupconsisting of H, hydroxy, halogen, a group containing a fatty acid, agroup containing a ketone body or pre-ketone body, a group containing anamino acid metabolite, an optionally substituted alkyl, an optionallysubstituted alkoxy, a monosaccharide, a sugar acid, phosphate, andsulfate; and

each of n and m is independently 0, 1, 2, 3, or 4.

In some embodiments, the compound includes at least one group containinga fatty acid. In particular embodiments, at least one R¹ is —OR^(A), inwhich R^(A) is a group containing a fatty acid, a group containing aketone body or pre-ketone body, or a group containing an amino acidmetabolite.

In particular embodiments,

is a single carbon-carbon bond. In certain embodiments, Q is —CH₂—.

In some embodiments, the acylated catechin polyphenol is of formula(I-a):

In certain embodiments, the acylated catechin polyphenol is of formula(I-b):

In particular embodiments, the acylated catechin polyphenol is offormula (I-c):

In further embodiments, the acylated catechin polyphenol is of formula(I-d):

In certain embodiments, the acylated catechin polyphenol is a compoundof formula (I-f):

In still further embodiments, n is 2. In certain embodiments, m is 1. Inparticular embodiments, m is 2. In some embodiments, m is 3. Inparticular embodiments, each R¹ is independently-OR^(A). In certainembodiments, each R³ is independently H or —OR^(A). In furtherembodiments, R² is H or —OR^(A). In yet further embodiments, each R^(A)is independently H, optionally substituted alkyl, a group containing afatty acid, a group containing a ketone body or pre-ketone body, or agroup containing an amino acid metabolite.

In some embodiments, R² is a group of formula:

where p is 1, 2, 3, or 4, and each R⁴ is independently selected from thegroup consisting of H, hydroxy, halogen, a group containing a fattyacid, a group containing a ketone body or pre-ketone body, a groupcontaining an amino acid metabolite, an optionally substituted alkyl, anoptionally substituted alkoxy, a monosaccharide, a sugar acid,phosphate, and sulfate.

In certain embodiments, p is 3. In particular embodiments, R⁴ isindependently H, hydroxy, halogen, a group containing a fatty acid, agroup containing a ketone body or pre-ketone body, a group containing anamino acid metabolite, or an optionally substituted alkoxy.

In some embodiments, the acylated catechin polyphenol is of formula(I-e):

In certain embodiments, each of R^(1A) and R^(1B) is independently—OR^(A). In particular embodiments, each of R^(3A), R^(3B), and R^(3C)is independently H, halogen, or —OR^(A).

In further embodiments, R² is a group of formula:

In yet further embodiments, R^(4A), R^(4B), and R^(4C) is independentlyH, hydroxy, halogen, a group containing a fatty acid, a group containinga ketone body or pre-ketone body, a group containing an amino acidmetabolite, or an optionally substituted alkoxy.

In some embodiments, each R^(A) is independently H, optionallysubstituted alkyl, fatty acid acyl, or optionally acylatedmonosaccharide.

In certain embodiments, the acylated catechin polyphenol includes atleast one fatty acid acyl (e.g., a short chain fatty acid acyl (e.g.,the short chain fatty acid acyl is acetyl, propionyl, or butyryl)).

In some embodiments, the acylated catechin polyphenol is compound 5.

Acylated Mesalamines

An acylated mesalamine of the invention may be a mesalamine, in whichone or more of —NH₂, —OH, or —COOH is replaced with an acyl, a groupcontaining a fatty acid, a group containing a ketone body or pre-ketonebody, or a group containing an amino acid metabolite. An acylatedmesalamine contains at least one group containing a fatty acid, groupcontaining a ketone body or pre-ketone body, or group containing anamino acid metabolite. A group containing a fatty acid, ketone body orpre-ketone body, or an amino acid metabolite is bonded to mesalaminethrough a glycosidic bond, ester bond, amide bond, carbonate linker, orcarbamate linker. In some embodiments, a group containing a fatty acidis bonded to mesalamine through a glycosidic bond. In some embodiments,acylated mesalamine is a compound of formula (II):

where

R¹ is H, alkyl, acyl, a group containing a fatty acid, a groupcontaining a ketone body or pre-ketone body, or a group containing anamino acid metabolite;

R² is H, alkyl, a group containing a fatty acid, a group containing aketone body or pre-ketone body, or a group containing an amino acidmetabolite; and

each R³ is independently H, alkyl, acyl, a group containing a fattyacid, a group containing a ketone body or pre-ketone body, or a groupcontaining an amino acid metabolite; or both R² groups combine to form:

The acylated mesalamine includes at least one group containing a fattyacid, at least one group containing a ketone body or pre-ketone body, orat least one group containing an amino acid metabolite.

In some embodiments, the acylated mesalamine is compound 44.

Acylated Shikimic Acid

An acylated active agent may be, e.g., an acylated shikimic acid of thefollowing structure:

or a salt thereof, where

each R¹ is independently H, acyl, alkyl, a group containing a fattyacid, a group containing a ketone body or pre-ketone body, or a groupcontaining an amino acid metabolite; and

R² is H, alkyl, a group containing a fatty acid, a group containing aketone body or pre-ketone body, or a group containing an amino acidmetabolite;

provided that the compound includes at least one group containing afatty acid, group containing a ketone body or pre-ketone body, or groupcontaining an amino acid metabolite.

Acylated Carotenoids

An acylated active agent may be, e.g., an acylated carotenoid. Acylatedcarotenoids are carotenoids, in which at least one hydroxyl group isreplaced with a substituent —OR, where each R is independently selectedfrom the group consisting of an acyl, alkyl, a group containing a fattyacid, a group containing a ketone body or pre-ketone body, and a groupcontaining an amino acid metabolite, provided that at least one R is agroup containing a fatty acid, a group containing a ketone body orpre-ketone body, or a group containing an amino acid metabolite.Non-limiting examples of an acylated carotenoid include astaxanthinhaving one or both hydroxyl groups independently substituted with anacyl, alkyl, group containing a fatty acid, or group containing a ketonebody or pre-ketone body, provided that at least one hydroxyl issubstituted with a group containing a fatty acid, a group containing aketone body or pre-ketone body, or a group containing an amino acidmetabolite

Acylated Ellagic Acid and Acylated Ellagic Acid Analogues

An acylated ellagic acid includes an ellagic acid core having one ormore hydroxyls substituted with an acyl (e.g., a fatty acid acyl). Anacylated ellagic acid analogue includes an ellagic acid analogue corehaving one or more hydroxyls substituted with an acyl (e.g., a fattyacid acyl).

An acylated ellagic acid is a compound of the following structures:

or a salt thereof,

where each R^(A) is independently H, alkyl, acyl, a group containing afatty acid, a group containing a ketone body or pre-ketone body, or agroup containing an amino acid metabolite; and each R^(B) isindependently H, alkyl, a group containing a fatty acid, a groupcontaining a ketone body or pre-ketone body, or a group containing anamino acid metabolite; provided that at least one R^(A) and/or at leastone R^(B), when present, is a group containing a fatty acid, a groupcontaining a ketone body or pre-ketone body, or a group containing anamino acid metabolite.

An acylated ellagic acid analogue is a compound of the followingstructure:

or a salt thereof,where

each of R², R³, and R⁴ is independently H or —OR^(A);

R⁶ is H or —(CO)—R^(5B);

R^(1A) is H or —OR^(A), and R^(5A) is —OH or —OR^(B); or R^(1A) andR^(5A) combine to form —O—;

R^(1B) is H or —OR^(A), and R^(5B) is absent, —OH, or —OR^(B); or R^(1B)and R^(SB) combine to form —O—;

each R^(A) is independently H, O-protecting group, alkyl, acyl, a groupcontaining a fatty acid, a group containing a ketone body or pre-ketonebody, or a group containing an amino acid metabolite;

each R^(B) is independently H, O-protecting group, alkyl, a groupcontaining a fatty acid, a group containing a ketone body or pre-ketonebody, or a group containing an amino acid metabolite; provided that atleast one R^(A) and/or at least one R^(B) is a group containing a fattyacid, a group containing a ketone body or pre-ketone body, or a groupcontaining an amino acid metabolite.

Non-limiting examples of ellagic acid analogues include urolithin A,urolithin B, urolithin C, urolithin D, urolithin E, and urolithin M5.

Acylated Hydroxybenzoic Add

An acylated active agent may be, e.g., an acylated hydroxybenzoic acidof the following structure:

or a salt thereof,

where

n is 1, 2, or 3;

each R¹ is independently H, acyl, alkyl, a group containing a fattyacid, a group containing a ketone body or pre-ketone body, or a groupcontaining an amino acid metabolite; and

R² is H, alkyl, a group containing a fatty acid, a group containing aketone body or pre-ketone body, or a group containing an amino acidmetabolite;

provided that the compound includes at least one group containing afatty acid, group containing a ketone body or pre-ketone body, or groupcontaining an amino acid metabolite.

Non-limiting examples of acylated hydroxybenzoic acids include salicylicacid, in which the phenolic hydroxyl is substituted with a groupcontaining a fatty acid, group containing a ketone body or pre-ketonebody, or group containing an amino acid metabolite; and gallic acid, inwhich one, two, or three phenolic hydroxyls are independentlysubstituted with groups containing a fatty acid, group containing aketone body or pre-ketone body, or group containing an amino acidmetabolite.

Acylated Sugar

An acylated active agent may be, e.g., an acylated sugar. An acylatedsugar is a monosaccharide having one or more hydroxyls substituted withalkyl, acyl, a group containing a fatty acid, a group containing aketone body or pre-ketone body, or a group containing an amino acidmetabolite. The monosaccharide is present in the pyranose or furanoseform. Preferably, the monosaccharide is present in the pyranose form.The monosaccharide may be an aldose or ketose. Non-limiting examples ofmonosaccharides are arabinose, xylose, fructose, galactose, glucose,ribose, tagatose, fucose, and rhamnose. In some embodiments, themonosaccharide is L-arabinose, D-xylose, fructose, galactose, D-glucose,D-ribose, D-tagatose, L-fucose, or L-rhamnose. Preferably, themonosaccharide is xylose, arabinose, rhamnose, fucose, glucosamine, ortagatose. The monosaccharide may include an anomeric carbon bonded to—OR, where R is H, alkyl, acyl, a group containing a fatty acid, a groupcontaining a ketone body or pre-ketone body, or a group containing anamino acid metabolite. Preferably, R is alkyl or a group containing afatty acid.

Methods

Acylated active agents described herein may be used to treat anautoimmune disorder in a subject in need thereof.

Without wishing to be bound by theory, metabolic products of themicrobiome can interact with the host immune system in several ways. Themetabolites can have effects remote to the gastrointestinal tract, forexample, through bidirectional interactions with the central nervoussystem. Examples include SCFA interacting with free fatty acidreceptors. Short-chain fatty acids may impact autoimmunity by expandingregulatory T cells and by suppressing the JNK1/P38 pathway. An acylatedactive agent described herein can biodegrade, for example, in the distalsmall intestine or colon, thereby providing high levels of the activeagent and fatty acids (e.g., short chain fatty acids) in the distal gut,where these compounds can interact with the immune system.

A method of treating an autoimmune disorder in a subject in need thereofmay include administering an acylated active agent (e.g., apharmaceutical or nutraceutical composition containing an acylatedactive agent) to the subject in need thereof.

In some embodiments, the components of the acylated active agent (e.g.,short chain fatty acid acyls (e.g., acetyl) and epigallocatechingallate) may act synergistically to treat an autoimmune disorder, e.g.,upon hydrolysis in the GI tract of the subject receiving the acylatedactive agent.

Non-limiting examples of autoimmune disorders include an inflammatorybowel disease, Addison's disease, alopecia areata, ankylosingspondylitis, antiphospholipid syndrome, hemolytic anemia, autoimmunehepatitis, Behcet's disease, Berger's disease, bullous pemphigoid,cardiomyopathy, celiac sprue, chronic fatigue immune dysfunctionsyndrome (CFIDS), chronic inflammatory demyelinating polyneuropathy,Churg-Strauss syndrome, cicatricial pemphigoid, cold agglutinin disease,type 1 diabetes, discoid lupus, essential mixed cryoglobulinemia,Graves' disease, Guillain-Barré syndrome, Hashimoto's thyroiditis,hypothyroidism, autoimmune lymphoproliferative syndrome (ALPS),idiopathic pulmonary fibrosis, idiopathic thrombocytopenia purpura(ITP), juvenile arthritis, lichen planus, lupus erythematosus, Meniere'sdisease, mixed connective tissue disease, multiple sclerosis, myastheniagravis, pemphigus vulgaris, pernicious anemia, polychondritis,autoimmune polyglandular syndromes, polymyalgia rheumatica,polymyositis, dermatomyositis, primary agammaglobulinemia, primarybiliary cirrhosis, psoriasis, psoriatic arthritis, Raynaud's phenomenon,Reiter's syndrome, rheumatic fever, rheumatoid arthritis, sarcoidosis,scleroderma, Sjögren's syndrome, stiff-man syndrome, Takayasu arteritis,giant cell arteritis, ulcerative colitis, uveitis, vasculitis, andgranulomatosis with polyangiitis. In some embodiments, the autoimmunedisorder is an inflammatory bowel disease (e.g., Crohn's disease orulcerative colitis).

Additionally or alternatively, acylated active agents described hereinmay be used for modulating an autoimmunity marker in a subject in needthereof.

A method of modulating an autoimmunity marker in a subject in needthereof may include administering an acylated active agent (e.g., apharmaceutical or nutraceutical composition containing an acylatedactive agent) to the subject in need thereof.

In some embodiments, the components of the acylated active agent (e.g.,short chain fatty acid acyls (e.g., acetyl) and epigallocatechingallate) may act synergistically to modulate an autoimmunity marker,e.g., upon hydrolysis in the GI tract of the subject receiving theacylated active agent.

Non-limiting examples of autoimmunity markers include markers for aninflammatory bowel disease, Addison's disease, alopecia areata,ankylosing spondylitis, antiphospholipid syndrome, hemolytic anemia,autoimmune hepatitis, Behcet's disease, Berger's disease, bullouspemphigoid, cardiomyopathy, celiac sprue, chronic fatigue immunedysfunction syndrome (CFIDS), chronic inflammatory demyelinatingpolyneuropathy, Churg-Strauss syndrome, cicatricial pemphigoid, coldagglutinin disease, type 1 diabetes, discoid lupus, essential mixedcryoglobulinemia, Graves' disease, Guillain-Barré syndrome, Hashimoto'sthyroiditis, hypothyroidism, autoimmune lymphoproliferative syndrome(ALPS), idiopathic pulmonary fibrosis, idiopathic thrombocytopeniapurpura (ITP), juvenile arthritis, lichen planus, lupus erythematosus,Meniere's disease, mixed connective tissue disease, multiple sclerosis,myasthenia gravis, pemphigus vulgaris, pernicious anemia,polychondritis, autoimmune polyglandular syndromes, polymyalgiarheumatica, polymyositis, dermatomyositis, primary agammaglobulinemia,primary biliary cirrhosis, psoriasis, psoriatic arthritis, Raynaud'sphenomenon, Reiter's syndrome, rheumatic fever, rheumatoid arthritis,sarcoidosis, scleroderma, Sjögren's syndrome, stiff-man syndrome,Takayasu arteritis, giant cell arteritis, ulcerative colitis, uveitis,vasculitis, and granulomatosis with polyangiitis. In some embodiments,the autoimmune marker is a marker for an inflammatory bowel disease(e.g., Crohn's disease or ulcerative colitis).

The autoimmunity markers include, for example, a CYP1A1 mRNA level,intestinal motility, mucus secretion, CD4⁺CD25⁺ Treg cell (e.g.,CD4⁺CD25⁺Foxp3⁺ Treg) count, T_(h)1 cell count, interleukin-8 (IL-8)level, macrophage inflammatory protein 1α (MIP-1α) level, macrophageinflammatory protein 1β (MIP-1β) level, NFκB level, inducible nitricoxide synthase (iNOS) level, matrix metallopeptidase 9 (MMP9) level,interferon γ (IFNγ) level, interleukin-17 (IL17) level, intercellularadhesion molecule (ICAM) level, CXCL13 level, 8-iso-prostaglandin F_(2α)(8-iso-PGF2α) level, IgA level, calprotectin level, lipocalin-2 level,short chain fatty acids level, and indoxyl sulfate level.

The autoimmunity markers can be measured in a sample from a subjectusing methods known in the art. For example, CD4⁺CD25⁺ Treg cell (e.g.,CD4⁺CD25⁺Foxp3⁺ Treg) count and Tn1 cell count are measured via routineblood test, followed by flow cytometry analysis of cell markers and/orcytokines (e.g., CD4, CD25, Foxp3, IFNγ, IL2, and/or IL4). NFκB and iNOSlevels can be measured using routine blood tests. Stool sample analysesmay be performed to measure an IgA level, calprotectin level,lipocalin-2 level, and short chain fatty acids level. Urine sampleanalysis may be performed to measure an indoxyl sulfate level. Mucussecretion can be assessed through biopsy or by analysis of fecal mattercontent. Mucus secretion can be measured using HT-29 cell counts or bymeasuring mucin gene expression in biopsy samples, e.g., by PCR (Recio,The impact of Food Bioactive on Health: In vitro and ex vivo models,Chapter 11, HT29 Cell line, (2015)). Intestinal motility can be assessedusing gastrointestinal scintigraphy (e.g., wireless pH and motilitycapsules) or by examining effect of a test article on its ability toimprove transepithelial electrical resistance (TEER) in either a cellline (e.g., CACO-2) or on a co-culture complex system (e.g., MATEKepi-intestinal) (Kickman, J. Lab. Autom., 20:107-126, 2015).Gastrointestinal permeability can be measured using a dual sugarabsorption test known in the art. For example, dual sugar absorptiontest involves administering a predetermined amount of a drink containinglactulose and mannitol, and measuring absorption of these two sugarsover six hours. Abdominal pain is typically assessed by a survey.Gastrointestinal bleeding may be assessed by the presence or absence ofblood in a stool sample from a subject. Gastrointestinal inflammationcan be assessed by biopsy.

In some embodiments, upon administration to a subject in need thereof,an acylated active agent described herein increases an autoimmunemarker, e.g., intestinal motility, CD4⁺CD25⁺ Treg cell count, shortchain fatty acids level, or mucus secretion in a subject (e.g., at least5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%,75%, 80%, 85%, 90%, 95% or 98% or more relative to prior toadministration). In some embodiments, upon administration to a subjectin need thereof, an acylated active agent described herein increases anautoimmune marker, e.g., a CYP1A1 mRNA level in a subject (e.g., atleast 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%,70%, 75%, 80%, 85%, 90%, 95% or 98% or more relative to prior toadministration). In certain embodiments, upon administration to asubject in need thereof, an acylated active agent described hereindecreases an autoimmune marker, e.g., iNOS, MMP9, IFNγ, IL17, ICAM,CXCL13, 8-iso-PGF2a in a subject (e.g., at least 5%, 10%, 15%, 20%, 25%,30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or98% or more relative to prior to administration). In certainembodiments, upon administration to a subject in need thereof, anacylated active agent described herein decreases an interleukin-8 (IL-8)level in a subject (e.g., at least 5%, 10%, 15%, 20%, 25%, 30%, 35%,40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or 98% ormore relative to prior to administration). In certain embodiments, uponadministration to a subject in need thereof, an acylated active agentdescribed herein decreases a macrophage inflammatory protein 1α (MIP-1α)level in a subject (e.g., at least 5%, 10%, 15%, 20%, 25%, 30%, 35%,40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or 98% ormore relative to prior to administration). In certain embodiments, uponadministration to a subject in need thereof, an acylated active agentdescribed herein decreases macrophage inflammatory protein 1β (MIP-1β)level in a subject (e.g., at least 5%, 10%, 15%, 20%, 25%, 30%, 35%,40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or 98% ormore relative to prior to administration). In further embodiments, uponadministration to a subject in need thereof, an acylated active agentdescribed herein modulates (increases or decreases) an autoimmunemarker, e.g., T_(h)1 cell count in a subject (e.g., at least 5%, 10%,15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%,85%, 90%, 95% or 98% or more relative to prior to administration). TheT_(h)1 cell count increase or decrease may be desirable depending on theparticular condition and its state. An attendant doctor or nursepractitioner can determine whether an increase or a decrease in theT_(h)1 cell count is desired.

In some embodiments, an acylated active agent described herein decreasesgastrointestinal inflammation (upper intestine, cecum, ileum, colon,rectum) in a subject (e.g., at least 5%, 10%, 15%, 20%, 25%, 30%, 35%,40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or 98% ormore relative to prior to administration)). In certain embodiments, anacylated active agent described herein decreases abdominal pain (e.g.,incidence and/or intensity) in a subject (e.g., at least 5%, 10%, 15%,20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%,90%, 95% or 98% or more relative to prior to administration). Inparticular embodiments, an acylated active agent described hereindecreases gastrointestinal permeability in a subject (e.g., at least 5%,10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%,80%, 85%, 90%, 95% or 98% or more relative to prior to administration).In further embodiments, an acylated active agent described hereinincreases intestinal motility or frequency of bowel movements in asubject (e.g., at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%,55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or 98% or more relative toprior to administration). In yet further embodiments, an acylated activeagent described herein decreases intestinal motility or frequency ofbowel movements in a subject (e.g., at least 5%, 10%, 15%, 20%, 25%,30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or98% or more relative to prior to administration). In still furtherembodiments, an acylated active agent described herein decreasesgastrointestinal bleeding in a subject (e.g., at least 5%, 10%, 15%,20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%,90%, 95% or 98% or more relative to prior to administration). In otherembodiments, am acylated active agent described herein decreases orincreases mucus secretion or improves mucosal health in agastrointestinal cell, tissue or in a subject (e.g., at least 5%, 10%,15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%,85%, 90%, 95% or 98% or more relative to prior to administration).

Pharmaceutical and Nutraceutical Compositions

The acylated active agents disclosed herein may be formulated intopharmaceutical or nutraceutical compositions for administration to humansubjects in a biologically compatible form suitable for administrationin vivo. Pharmaceutical and nutraceutical compositions typically includean acylated active agent as described herein and a physiologicallyacceptable excipient (e.g., a pharmaceutically acceptable excipient).

The acylated active agents described herein can also be used in the formof the free acid/base, in the form of salts, zwitterions, or assolvates. All forms are within the scope of the invention. The acylatedactive agents, salts, zwitterions, solvates, or pharmaceutical ornutraceutical compositions thereof, may be administered to a subject ina variety of forms depending on the selected route of administration, aswill be understood by those skilled in the art. The acylated activeagents described herein may be administered, for example, by oral,parenteral, buccal, sublingual, nasal, rectal, patch, pump, ortransdermal administration, and the pharmaceutical or nutraceuticalcompositions formulated accordingly. Parenteral administration includesintravenous, intraperitoneal, subcutaneous, intramuscular,transepithelial, nasal, intrapulmonary, intrathecal, rectal, and topicalmodes of administration. Parenteral administration may be by continuousinfusion over a selected period of time.

For human use, an acylated active agent disclosed herein can beadministered alone or in admixture with a pharmaceutical ornutraceutical carrier selected regarding the intended route ofadministration and standard pharmaceutical practice. Pharmaceutical andnutraceutical compositions for use in accordance with the presentinvention thus can be formulated in a conventional manner using one ormore physiologically acceptable carriers comprising excipients andauxiliaries that facilitate processing of acylated active agentsdisclosed herein into preparations which can be used pharmaceutically.

This disclosure also includes pharmaceutical and nutraceuticalcompositions which can contain one or more physiologically acceptablecarriers. In making the pharmaceutical or nutraceutical compositions ofthe invention, the active ingredient is typically mixed with anexcipient, diluted by an excipient or enclosed within such a carrier inthe form of, for example, a capsule, sachet, paper, or other container.When the excipient serves as a diluent, it can be a solid, semisolid, orliquid material (e.g., normal saline), which acts as a vehicle, carrieror medium for the active ingredient. Thus, the compositions can be inthe form of tablets, powders, lozenges, sachets, cachets, elixirs,suspensions, emulsions, solutions, syrups, and soft and hard gelatincapsules. As is known in the art, the type of diluent can vary dependingupon the intended route of administration. The resulting compositionscan include additional agents, e.g., preservatives. Nutraceuticalcompositions may be administered enterally (e.g., orally). Anutraceutical composition may be a nutraceutical oral formulation (e.g.,a tablet, powder, lozenge, sachet, cachet, elixir, suspension, emulsion,solution, syrup, or soft or hard gelatin capsule), food additive (e.g.,a food additive as defined in 21 C.F.R. § 170.3), food product (e.g.,food for special dietary use as defined in 21 C.F.R. § 105.3), ordietary supplement (e.g., where the active agent is a dietary ingredient(e.g., as defined in 21 U.S.C. § 321 (ff))). Active agents can be usedin nutraceutical applications and as food additive or food products.Non-limiting examples of compositions including an active agent of theinvention are a bar, drink, shake, powder, additive, gel, or chew.

The excipient or carrier is selected on the basis of the mode and routeof administration. Suitable pharmaceutical carriers, as well aspharmaceutical necessities for use in pharmaceutical formulations, aredescribed in Remington: The Science and Practice of Pharmacy, 21^(st)Ed., Gennaro, Ed., Lippencott Williams & Wilkins (2005), a well-knownreference text in this field, and in the USP/NF (United StatesPharmacopeia and the National Formulary). Examples of suitableexcipients are lactose, dextrose, sucrose, sorbitol, mannitol, starches,gum acacia, calcium phosphate, alginates, tragacanth, gelatin, calciumsilicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose,water, syrup, and methyl cellulose. The formulations can additionallyinclude: lubricating agents, e.g., talc, magnesium stearate, and mineraloil; wetting agents; emulsifying and suspending agents; preservingagents, e.g., methyl- and propylhydroxy-benzoates; sweetening agents;and flavoring agents. Other exemplary excipients are described inHandbook of Pharmaceutical Excipients, 6^(th) Edition, Rowe et al.,Eds., Pharmaceutical Press (2009).

These pharmaceutical and nutraceutical compositions can be manufacturedin a conventional manner, e.g., by conventional mixing, dissolving,granulating, dragee-making, levigating, emulsifying, encapsulating,entrapping, or lyophilizing processes. Methods well known in the art formaking formulations are found, for example, in Remington: The Scienceand Practice of Pharmacy, 21^(st) Ed., Gennaro, Ed., Lippencott Williams& Wilkins (2005), and Encyclopedia of Pharmaceutical Technology, eds. J.Swarbrick and J. C. Boylan, 1988-1999, Marcel Dekker, New York. Properformulation is dependent upon the route of administration chosen. Theformulation and preparation of such compositions is well-known to thoseskilled in the art of pharmaceutical and nutraceutical formulation. Inpreparing a formulation, the acylated active agents can be milled toprovide the appropriate particle size prior to combining with the otheringredients. If the acylated active agent is substantially insoluble, itcan be milled to a particle size of less than 200 mesh. If the acylatedactive agent is substantially water soluble, the particle size can beadjusted by milling to provide a substantially uniform distribution inthe formulation, e.g., about 40 mesh.

Dosages

The dosage of the acylated active agent used in the methods describedherein, or pharmaceutically acceptable salts or prodrugs thereof, orpharmaceutical or nutraceutical compositions thereof, can vary dependingon many factors, e.g., the pharmacodynamic properties of the acylatedactive agent; the mode of administration; the age, health, and weight ofthe recipient; the nature and extent of the symptoms; the frequency ofthe treatment, and the type of concurrent treatment, if any; and theclearance rate of the acylated active agent in the subject to betreated. One of skill in the art can determine the appropriate dosagebased on the above factors. The acylated active agents used in themethods described herein may be administered initially in a suitabledosage that may be adjusted as required, depending on the clinicalresponse. In general, a suitable daily dose of an acylated active agentdisclosed herein will be that amount of the acylated active agent thatis the lowest dose effective to produce a therapeutic effect. Such aneffective dose will generally depend upon the factors described above.

An acylated active agent disclosed herein may be administered to thesubject in a single dose or in multiple doses. When multiple doses areadministered, the doses may be separated from one another by, forexample, 1-24 hours, 1-7 days, or 1-4 weeks. The acylated active agentmay be administered according to a schedule, or the acylated activeagent may be administered without a predetermined schedule. It is to beunderstood that, for any particular subject, specific dosage regimesshould be adjusted overtime according to the individual need and theprofessional judgment of the person administering or supervising theadministration of the compositions.

The acylated active agents may be provided in a dosage form. In someembodiments, the unit dosage form may be an oral unit dosage form (e.g.,a tablet, capsule, suspension, liquid solution, powder, crystals,lozenge, sachet, cachet, elixir, syrup, and the like) or a food productserving (e.g., the active agents may be included as food additives ordietary ingredients). In certain embodiments, the dosage form isdesigned for administration of at least one acylated active agentdisclosed herein, where the total amount of an administered acylatedactive agent is from 0.1 g to 10 g (e.g., 0.5 g to 9 g, 0.5 g to 8 g,0.5 g to 7 g, 0.5 g to 6 g, 0.5 g to 5 g, 0.5 g to 1 g, 0.5 g to 1.5 g,0.5 g to 2 g, 0.5 g to 2.5 g, 1 g to 1.5 g, 1 g to 2 g, 1 g to 2.5 g,1.5 g to 2 g, 1.5 g to 2.5 g, or 2 g to 2.5 g). In other embodiments,the acylated active agent is consumed at a rate of 0.1 g to 10 g per day(e.g., 0.5 g to 9 g, 0.5 g to 8 g, 0.5 g to 7 g, 0.5 g to 6 g, 0.5 g to5 g, 0.5 g to 1 g per day, 0.5 g to 1.5 g per day, 0.5 g to 2 g per day,0.5 g to 2.5 g per day, 1 g to 1.5 g per day, 1 g to 2 g per day, 1 g to2.5 g per day, 1.5 g to 2 g per day, 1.5 g to 2.5 g per day, or 2 g to2.5 g per day) or more. The attending physician ultimately will decidethe appropriate amount and dosage regimen, an effective amount of theacylated active agent disclosed herein may be, for example, a totaldaily dosage of, e.g., between 0.5 g and 5 g (e.g., 0.5 to 2.5 g) of anyof the acylated active agent described herein. Alternatively, the dosageamount can be calculated using the body weight of the subject.Preferably, when daily dosages exceed 5 g/day, the dosage of theacylated active agent may be divided across two or three dailyadministration events.

In the methods of the invention, the time period during which multipledoses of an acylated active agent disclosed herein are administered to asubject can vary. For example, in some embodiments doses of the acylatedactive agents are administered to a subject over a time period that is1-7 days; 1-12 weeks; or 1-3 months. In other embodiments, the acylatedactive agents are administered to the subject over a time period thatis, for example, 4-11 months or 1-30 years. In yet other embodiments,the acylated active agents disclosed herein are administered to asubject at the onset of symptoms. In any of these embodiments, theamount of the acylated active agent that is administered may vary duringthe time period of administration. When an acylated active agent isadministered daily, administration may occur, for example, 1, 2, 3, or 4times per day.

Formulations

An acylated active agent described herein may be administered to asubject with a pharmaceutically acceptable diluent, carrier, orexcipient, in unit dosage form. Conventional pharmaceutical practice maybe employed to provide suitable formulations or compositions toadminister the acylated active agent to subjects suffering from adisorder. Administration may begin before the subject is symptomatic.

Exemplary routes of administration of the acylated active agentsdisclosed herein or pharmaceutical or nutraceutical compositionsthereof, used in the present invention include oral, sublingual, buccal,transdermal, intradermal, intramuscular, parenteral, intravenous,intra-arterial, intracranial, subcutaneous, intraorbital,intraventricular, intraspinal, intraperitoneal, intranasal, inhalation,and topical administration. The acylated active agents desirably areadministered with a physiologically acceptable carrier (e.g., apharmaceutically acceptable carrier). Pharmaceutical formulations of theacylated active agents described herein formulated for treatment of thedisorders described herein are also part of the present invention. Insome preferred embodiments, the acylated active agents disclosed hereinare administered to a subject orally. In other preferred embodiments,the acylated active agents disclosed herein are administered to asubject topically.

Formulations for Oral Administration

The pharmaceutical and nutraceutical compositions contemplated by theinvention include those formulated for oral administration (“oral dosageforms”). Oral dosage forms can be, for example, in the form of tablets,capsules, a liquid solution or suspension, a powder, or liquid or solidcrystals, which contain the active ingredient(s) in a mixture withphysiologically acceptable excipients (e.g., pharmaceutically acceptableexcipients). These excipients may be, for example, inert diluents orfillers (e.g., sucrose, sorbitol, sugar, mannitol, microcrystallinecellulose, starches including potato starch, calcium carbonate, sodiumchloride, lactose, calcium phosphate, calcium sulfate, or sodiumphosphate); granulating and disintegrating agents (e.g., cellulosederivatives including microcrystalline cellulose, starches includingpotato starch, croscarmellose sodium, alginates, oralginic acid);binding agents (e.g., sucrose, glucose, sorbitol, acacia, alginic acid,sodium alginate, gelatin, starch, pregelatinized starch,microcrystalline cellulose, magnesium aluminum silicate,carboxymethylcellulose sodium, methylcellulose, hydroxypropylmethylcellulose, ethylcellulose, polyvinylpyrrolidone, or polyethyleneglycol); and lubricating agents, glidants, and antiadhesives (e.g.,magnesium stearate, zinc stearate, stearic acid, silicas, hydrogenatedvegetable oils, or talc). Other physiologically acceptable excipients(e.g., pharmaceutically acceptable excipients) can be colorants,flavoring agents, plasticizers, humectants, buffering agents, and thelike.

Formulations for oral administration may also be presented as chewabletablets, as hard gelatin capsules where the active ingredient is mixedwith an inert solid diluent (e.g., potato starch, lactose,microcrystalline cellulose, calcium carbonate, calcium phosphate orkaolin), or as soft gelatin capsules where the active ingredient ismixed with water or an oil medium, for example, peanut oil, liquidparaffin, or olive oil. Powders, granulates, and pellets may be preparedusing the ingredients mentioned above under tablets and capsules in aconventional manner using, e.g., a mixer, a fluid bed apparatus or aspray drying equipment.

Controlled release compositions for oral use may be constructed torelease the active drug by controlling the dissolution and/or thediffusion of the active drug substance. Any of a number of strategiescan be pursued in order to obtain controlled release and the targetedplasma concentration versus time profile. In one example, controlledrelease is obtained by appropriate selection of various formulationparameters and ingredients, including, e.g., various types of controlledrelease compositions and coatings. Examples include single or multipleunit tablet or capsule compositions, oil solutions, suspensions,emulsions, microcapsules, microspheres, nanoparticles, patches, andliposomes. In certain embodiments, compositions include biodegradable,pH, and/or temperature-sensitive polymer coatings.

Dissolution or diffusion controlled release can be achieved byappropriate coating of a tablet, capsule, pellet, or granulateformulation of acylated active agents, or by incorporating the acylatedactive agent into an appropriate matrix. A controlled release coatingmay include one or more of the coating substances mentioned aboveand/or, e.g., shellac, beeswax, glycowax, castor wax, carnauba wax,stearyl alcohol, glyceryl monostearate, glyceryl distearate, glycerolpalmitostearate, ethylcellulose, acrylic resins, dl-polylactic acid,cellulose acetate butyrate, polyvinyl chloride, polyvinyl acetate, vinylpyrrolidone, polyethylene, polymethacrylate, methylmethacrylate,2-hydroxymethacrylate, methacrylate hydrogels, 1,3 butylene glycol,ethylene glycol methacrylate, and/or polyethylene glycols. In acontrolled release matrix formulation, the matrix material may alsoinclude, e.g., hydrated methylcellulose, carnauba wax and stearylalcohol, carbopol 934, silicone, glyceryl tristearate, methylacrylate-methyl methacrylate, polyvinyl chloride, polyethylene, and/orhalogenated fluorocarbon.

The liquid forms in which the acylated active agents and compositions ofthe present invention can be incorporated for administration orallyinclude aqueous solutions, suitably flavored syrups, aqueous or oilsuspensions, and flavored emulsions with edible oils, e.g., cottonseedoil, sesame oil, coconut oil, or peanut oil, as well as elixirs andsimilar pharmaceutical and nutraceutical vehicles.

Formulations for Buccal Administration

Dosages for buccal or sublingual administration typically are 0.1 to 500mg per single dose as required. In practice, the physician determinesthe actual dosing regimen which is most suitable for an individualsubject, and the dosage varies with the age, weight, and response of theparticular subject. The above dosages are exemplary of the average case,but individual instances exist where higher or lower dosages aremerited, and such are within the scope of this invention.

For buccal administration, the compositions may take the form oftablets, lozenges, etc. formulated in a conventional manner. Liquid drugformulations suitable for use with nebulizers and liquid spray devicesand electrohydrodynamic (EHD) aerosol devices will typically include aacylated active agent disclosed herein with a pharmaceuticallyacceptable carrier. Preferably, the pharmaceutically acceptable carrieris a liquid, e.g., alcohol, water, polyethylene glycol, or aperfluorocarbon. Optionally, another material may be added to alter theaerosol properties of the solution or suspension of acylated activeagents disclosed herein. Desirably, this material is liquid, e.g., analcohol, glycol, polyglycol, or a fatty acid. Other methods offormulating liquid drug solutions or suspension suitable for use inaerosol devices are known to those of skill in the art (see, e.g., U.S.Pat. Nos. 5,112,598 and 5,556,611, each of which is herein incorporatedby reference).

Formulations for Nasal or Inhalation Administration

The acylated active agents may also be formulated for nasaladministration. Compositions for nasal administration also mayconveniently be formulated as aerosols, drops, gels, and powders. Theformulations may be provided in a single or multidose form. In the caseof a dropper or pipette, dosing may be achieved by the subjectadministering an appropriate, predetermined volume of the solution orsuspension. In the case of a spray, this may be achieved, for example,by means of a metering atomizing spray pump.

The acylated active agents may further be formulated for aerosoladministration, particularly to the respiratory tract by inhalation andincluding intranasal administration. The acylated active agents fornasal or inhalation administration will generally have a small particlesize for example on the order of five (5) microns or less. Such aparticle size may be obtained by means known in the art, for example bymicronization. The active ingredient is provided in a pressurized packwith a suitable propellant, e.g., a chlorofluorocarbon (CFC), forexample, dichlorodifluoromethane, trichlorofluoromethane, ordichlorotetrafluoroethane, or carbon dioxide, or other suitable gas. Theaerosol may conveniently also contain a surfactant, e.g., lecithin. Thedose of drug may be controlled by a metered valve. Alternatively, theactive ingredients may be provided in a form of a dry powder, e.g., apowder mix of the acylated active agent in a suitable powder base, e.g.,lactose, starch, and starch derivatives, e.g., hydroxypropylmethylcellulose, and polyvinylpyrrolidine (PVP). The powder carrier will forma gel in the nasal cavity. The powder composition may be presented inunit dose form for example in capsules or cartridges of e.g., gelatin orblister packs from which the powder may be administered by means of aninhaler.

Aerosol formulations typically include a solution or fine suspension ofthe active substance in a physiologically acceptable aqueous ornon-aqueous solvent and are usually presented in single or multidosequantities in sterile form in a sealed container, which can take theform of a cartridge or refill for use with an atomizing device.Alternatively, the sealed container may be a unitary dispensing device,e.g., a single dose nasal inhaler or an aerosol dispenser fitted with ametering valve which is intended for disposal after use. Where thedosage form comprises an aerosol dispenser, it will contain apropellant, which can be a compressed gas, e.g., compressed air or anorganic propellant, e.g., fluorochlorohydrocarbon. The aerosol dosageforms can also take the form of a pump-atomizer.

Formulations for Parenteral Administration

The acylated active agents described herein for use in the methods ofthe invention can be administered in a pharmaceutically acceptableparenteral (e.g., intravenous or intramuscular) formulation as describedherein. The pharmaceutical formulation may also be administeredparenterally (intravenous, intramuscular, subcutaneous or the like) indosage forms or formulations containing conventional, non-toxicpharmaceutically acceptable carriers and adjuvants. In particular,formulations suitable for parenteral administration include aqueous andnon-aqueous sterile injection solutions which may contain anti-oxidants,buffers, bacteriostats, and solutes which render the formulationisotonic with the blood of the intended recipient; and aqueous andnon-aqueous sterile suspensions which may include suspending agents andthickening agents. For example, to prepare such a composition, theacylated active agents disclosed herein may be dissolved or suspended ina parenterally acceptable liquid vehicle.

Among acceptable vehicles and solvents that may be employed are water,water adjusted to a suitable pH by addition of an appropriate amount ofhydrochloric acid, sodium hydroxide or a suitable buffer,1,3-butanediol, Ringer's solution and isotonic sodium chloride solution.The aqueous formulation may also contain one or more preservatives, forexample, methyl, ethyl or n-propyl p-hydroxybenzoate. Additionalinformation regarding parenteral formulations can be found, for example,in the United States Pharmacopeia-National Formulary (USP-NF), hereinincorporated by reference.

The parenteral formulation can be any of the five general types ofpreparations identified by the USP-NF as suitable for parenteraladministration:

-   -   (1) “Drug Injection:” a liquid preparation that is a drug        substance (e.g., an acylated active agent disclosed herein or a        solution thereof);    -   (2) “Drug for Injection:” the drug substance (e.g., an acylated        active agent disclosed herein) as a dry solid that will be        combined with the appropriate sterile vehicle for parenteral        administration as a drug injection;    -   (3) “Drug Injectable Emulsion:” a liquid preparation of the drug        substance (e.g., an acylated active agent disclosed herein) that        is dissolved or dispersed in a suitable emulsion medium;    -   (4) “Drug Injectable Suspension:” a liquid preparation of the        drug substance (e.g., an acylated active agent disclosed herein)        suspended in a suitable liquid medium; and    -   (5) “Drug for Injectable Suspension:” the drug substance (e.g.,        an acylated active agent disclosed herein) as a dry solid that        will be combined with the appropriate sterile vehicle for        parenteral administration as a drug injectable suspension.

Exemplary formulations for parenteral administration include solutionsof the acylated active agents prepared in water suitably mixed with asurfactant, e.g., hydroxypropylcellulose. Dispersions can also beprepared in glycerol, liquid polyethylene glycols, DMSO and mixturesthereof with or without alcohol, and in oils. Under ordinary conditionsof storage and use, these preparations may contain a preservative toprevent the growth of microorganisms. Conventional procedures andingredients for the selection and preparation of suitable formulationsare described, for example, in Remington: The Science and Practice ofPharmacy, 21^(st) Ed., Gennaro, Ed., Lippencott Williams & Wlkins (2005)and in The United States Pharmacopeia: The National Formulary (USP 36NF31), published in 2013.

Formulations for parenteral administration may, for example, containexcipients, sterile water, or saline, polyalkylene glycols, e.g.,polyethylene glycol, oils of vegetable origin, or hydrogenatednapthalenes. Biocompatible, biodegradable lactide polymer,lactide/glycolide copolymer, or polyoxyethylene-polyoxypropylenecopolymers may be used to control the release of the acylated activeagents or biologically active agents within acylated active agents.Other potentially useful parenteral delivery systems for acylated activeagents include ethylene-vinyl acetate copolymer particles, osmoticpumps, implantable infusion systems, and liposomes. Formulations forinhalation may contain excipients, for example, lactose, or may beaqueous solutions containing, for example, polyoxyethylene-9-laurylether, glycocholate and deoxycholate, or may be oily solutions foradministration in the form of nasal drops, or as a gel.

The parenteral formulation can be formulated for prompt releaseorforsustained/extended release of the acylated active agent. Exemplaryformulations for parenteral release of the acylated active agentinclude: aqueous solutions, powders for reconstitution, cosolventsolutions, oil/water emulsions, suspensions, oil-based solutions,liposomes, microspheres, and polymeric gels.

Preparation of Acylated Active Agents

Acylated active agents may be prepared using synthetic methods andreaction conditions known in the art. Optimum reaction conditions andreaction times may vary depending on the reactants used. Unlessotherwise specified, solvents, temperatures, pressures, and otherreaction conditions may be selected by one of ordinary skill in the art.

Ester Preparation Strategy #1 (Acylation)

In Scheme 1, a polyphenolic compound, compound 1 where n represents aninteger from 1 to 15, is treated with an acylating agent, compound 2, inan appropriate solvent, optionally in the presence of a catalyst.Suitable catalysts include pyridine, dimethylaminopyridine,trimethylamine and the like. The catalyst can be used in quantitiesranging from 0.01 to 1.1 equivalents, relative to compound 2. Suitablesolvents include methylene chloride, ethyl acetate, diethyl ether,tetrahydrofuran, 1,4-dioxane, 1,2-dimethoxyethane, toluene, combinationsthereof and the like. Reaction temperatures range from −10° C. to theboiling point of the solvent used; reaction completion times range from1 to 96 h. Suitable acylating agents include acyl chlorides, acylfluorides, acyl bromides, carboxylic acid anhydrides whether symmetricalor not. A suitable acylating agent may also be generated in situ byprior reaction of a carboxylic acid with an activating reagent such asEDC or EEDQ or the like. The acylating agents can be used in quantitiesranging from 0.5 to 15 equivalents relative to compound 1.

The product, compound 3, can be purified by methods known to those ofskill in the art.

Ester Preparation Strategy #2 (Acylation)

In some cases, the polyphenolic compound 1 may contain a functionalgroup, Y, required to remain unreacted in the course of ester formation.In this case, it is appropriate to protect the functional group, Y, inthe polyphenolic compound from acylation. This functional group may bean amino group or a hydroxyl group or other functionality with a labilehydrogen attached to a heteroatom. Such polyphenol esters can beprepared according to Scheme 2.

In Scheme 2 Step 1, compound 1, a polyphenolic compound containing afunctional group Y with a labile hydrogen in need of protection, istreated with a protecting reagent such as BOC anhydride,benzyoxycarbonyl chloride, FMOC chloride, benzyl bromide and the like inan appropriate solvent, optionally in the presence of a catalyst toprovide compound 2 scheme 2. Compound 2 can be purified by methods knownto those of skill in the art.

In Scheme 2 Step 2, compound 2 is treated with an acylating agent,compound 3, in an appropriate solvent, optionally in the presence of acatalyst. Suitable catalysts include pyridine, dimethylaminopyridine,trimethylamine and the like. The catalyst can be used in quantitiesranging from 0.01 to 1.1 equivalents, relative to compound 2. Suitablesolvents include methylene chloride, ethyl acetate, diethyl ether,tetrahydrofuran, 1,4-dioxane, 1,2-dimethoxyethane, toluene, combinationsthereof and the like. Reaction temperatures range from −10° C. to theboiling point of the solvent used; reaction completion times range from1 to 96 h. Suitable acylating agents include acyl chlorides, acylfluorides, acyl bromides, carboxylic acid anhydrides whether symmetricalor not. A suitable acylating agent may also be generated in situ byprior reaction of a carboxylic acid with an activating reagent such asEDC or EEDQ or the like. The acylating agents can be used in quantitiesranging from 0.5 to 15 equivalents, relative to compound 3. Compound 4can be purified by methods known to those of skill in the art.

In Scheme 2 Step 3, compound 4 is subjected to conditions that cleavethe protecting group, PG.

In the case of a BOC protecting group, the protecting group of compound4 is removed under acidic conditions to give compound 5 of theinvention. Suitable acids include trifluoroacetic acid, hydrochloricacid, p-toluenesulfonic acid and the like.

In the case of an FMOC protecting group, the protecting group ofcompound 4 is removed under basic conditions to give compound 5 of theinvention. Suitable bases include piperidine, triethylamine and thelike. Suitable solvents include DMF, NMP dichoromethane and the like.The FMOC group is also removed under non-basic conditions such as bytreatment with tetrabutylammonium fluoride trihydrate in a suitablesolvent such as DMF. The FMOC group is also removed by catalytichydrogenation. Suitable catalysts for hydrogenation include 10%palladium-on-charcoal and palladium (II) acetate and the like. Suitablesolvents for hydrogenation include DMF, ethanol, and the like

In the case of a benzyloxycarbonyl or benzyl protecting group theprotecting group of compound 4 is removed by hydrogenation to givecompound 5. Suitable catalysts for hydrogenation include 10%Palladium-on-charcoal and Palladium acetate and the like. Suitablesolvents for hydrogenation include DMF, ethanol, methanol, ethylacetate, and the like. The product, compound 5, can be purified bymethods known to those of skill in the art.

Ester Preparation Strategy #3 (Acylation)

In Scheme 3 Step 1, compound 1, an acyl compound containing a functionalgroup Y with a labile hydrogen in need on protection, is treated with aprotecting reagent such as BOC anhydride, benzyoxycarbonyl chloride,FMOC chloride, benzyl bromide and the like in an appropriate solvent,optionally in the presence of a catalyst to provide compound 2 scheme 3.Compound 2 can be purified by methods known to those of skill in theart.

In Scheme 3 Step 2, compound 2 is treated with an activating reagentsuch as thionyl chloride, phosphorus oxychloride, EDC or EEDQ or thelike to generate the activated acyl compound 3.

In Scheme 3 Step 3, the polyphenol compound 4 is treated with theactivated acyl compound 3, in an appropriate solvent, optionally in thepresence of a catalyst. Suitable catalysts include pyridine,dimethylaminopyridine, trimethylamine and the like to generate compound5. The catalyst can be used in quantities ranging from 0.01 to 1.1equivalents, relative to compound 3. Suitable solvents include methylenechloride, ethyl acetate, diethyl ether, tetrahydrofuran, 1,4-dioxane,1,2-dimethoxyethane, toluene, combinations thereof and the like.Reaction temperatures range from −10° C. to the boiling point of thesolvent used; reaction completion times range from 1 to 96 h. Theactivated acyl compound 3 can be used in quantities ranging from 0.5 to15 equivalents relative to compound 4.

In Scheme 3 Step 4, compound 5 is subjected to conditions designed tocleave the protecting group, PG, illustrated in Scheme 2 above. Theproduct, compound 6, can be purified by methods known to those of skillin the art.

Ester Preparation Strategy #4 (Acylation)

In Scheme 4 Step 1a poly-ol compound, compound 1, where R represents anon-aromatic cyclic or acyclic moiety and n represents an integer from 1to 15, is treated with an acylating agent, compound 2, in an appropriatesolvent, optionally in the presence of a catalyst. Suitable catalystsinclude pyridine, dimethylaminopyridine, trimethylamine and the like.The catalyst can be used in quantities ranging from 0.01 to 1.1equivalents, relative to compound 2. Suitable solvents include methylenechloride, ethyl acetate, diethyl ether, tetrahydrofuran, 1,4-dioxane,1,2-dimethoxyethane, toluene, combinations thereof and the like.Reaction temperatures range from −10° C. to the boiling point of thesolvent used; reaction completion times range from 1 to 96 h. Suitableacylating agents include acyl chlorides, acyl fluorides, acyl bromides,carboxylic acid anhydrides whether symmetrical or not. A suitableacylating agent may also be generated in situ by prior reaction of acarboxylic acid with an activating reagent such as EDC or EEDQ or thelike. The acylating agents can be used in quantities ranging from 0.5 to15 equivalents, relative to compound 1. The product, compound 3, can bepurified by methods known to those of skill in the art.

Ester Preparation Strategy #5 (Baeyer-Villiger Oxidation)

In Scheme 5 Step 1, a ketone compound, compound 1, where R and R1represent non-aromatic cyclic or acyclic moieties, is treated with aperoxide or peroxyacid agent, such as meta-chloroperbenzoic acid,performic acid, peracetic acid, hydrogen peroxide, tert-butylhydroperoxide and the like in an appropriate solvent, optionally in thepresence of a catalyst. Suitable solvents include methylene chloride,diethyl ether, combinations thereof and the like. Suitable catalystsinclude BF3, carboxylic acids and the like. Reaction temperatures rangefrom −10° C. to the boiling point of the solvent used; reactioncompletion times range from 1 to 96 h. The product, compound 2, can bepurified by methods known to those of skill in the art.

The R and R1 groups of compound 1 in Scheme 5 may optionally includeadditional ketone functionality that can undergo reaction. In additionthe R and R1 groups of compound 1 may form a ring.

Ester Preparation Strategy #6 (Mitsunobu Reaction)

In Scheme 6 Step 1, a mixture of an alcohol compound, compound 1, whereR represents a non-aromatic cyclic or acyclic moiety, and a carboxylicacid, compound 2 where R1 represents an alkanoyl group optionallysubstituted with one or more protected hydroxyl groups or oxo is treatedwith triphenylphosphine and a diazo compound such asdiethylazodicarboxylate (DEAD) and the like in an appropriate solvent.Suitable solvents include methylene chloride, THF, acetonitrile,toluene, diethyl ether, combinations thereof and the like. Reactiontemperatures range from −10° C. to the boiling point of the solventused; reaction completion times range from 1 to 96 h. The product,compound 3 can be purified by methods known to those of skill in theart.

Where compound 3 is optionally substituted by one or more protectedalcohol groups deprotection is accomplished by the methods illustratedin Scheme 2 above.

Ester preparation strategy #7 (Nucleophilic Alkylation)

In Scheme 7 Step 1, a chloroformate compound, compound 1, where Rrepresents an aromatic moiety or a non-aromatic cyclic or acyclicmoiety, is treated, in an appropriate solvent, with an organometalliccompound, compound 2 where R1 represents an alkyl group optionallysubstituted with one or more protected hydroxyl groups and X representsa metal such as Cu, Zn, Mg which is optionally coordinated by one ormore counterions, such as chloride. Suitable solvents include methylenechloride, THF, acetonitrile, toluene, diethyl ether, combinationsthereof, and the like. Reaction temperatures range from −10° C. to theboiling point of the solvent used; reaction completion times range from1 to 96 h. The product, compound 3, can be purified by methods known tothose of skill in the art.

Compound 1 can be prepared from the corresponding alcohol or polyolcompounds by standard methods familiar to one skilled in the art.

Where compound 2 is optionally substituted by one or more protectedalcohol groups deprotection is accomplished by the methods illustratedin Scheme 2 above.

Further modification of the initial product by methods known in the artand illustrated in the examples below, may be used to prepare additionalcompounds of this invention.

Ester Preparation Strategy #8 (Acylation)

In Scheme 8 Step 1, compound 1, an acyl compound containing a hydroxylgroup to be acylated, is treated with a protecting reagent such asbenzyl bromide and the like in an appropriate solvent, optionally in thepresence of a catalyst to provide compound 2 scheme 8. Compound 2 can bepurified by methods known to those of skill in the art.

In scheme 8 Step 2, compound 2 is treated with an acylating agent, in anappropriate solvent, optionally in the presence of a catalyst. Suitablecatalysts include pyridine, dimethylaminopyridine, trimethylamine andthe like. The catalyst can be used in quantities ranging from 0.01 to1.1 equivalents, relative to compound 2. Suitable solvents includemethylene chloride, ethyl acetate, diethyl ether, tetrahydrofuran,1,4-dioxane, 1,2-dimethoxyethane, toluene, combinations thereof and thelike. Reaction temperatures range from −10° C. to the boiling point ofthe solvent used; reaction completion times range from 1 to 96 h.Suitable acylating agents include acyl chlorides, acyl fluorides, acylbromides, carboxylic acid anhydrides whether symmetrical or not. Asuitable acylating agent may also be generated in situ by a reaction ofa carboxylic acid with an activating reagent such as EDC or EEDQ or thelike. The acylating agents can be used in quantities ranging from 0.5 to15 equivalents relative to compound 1.

In Scheme 8 Step 3, compound 3 is subjected to conditions that cleavethe protecting group, PG. In the case of a benzyl protecting group, theprotecting group of compound 3 is removed by hydrogenation to givecompound 4. Suitable catalysts for hydrogenation include 10%palladium-on-charcoal and palladium acetate and the like. Suitablesolvents for hydrogenation include, DMF, ethanol, methanol, ethylacetate and the like. The product, compound 4, can be purified bymethods known to those of skill in the art.

In Scheme 8 Step 4, compound 4 is treated with an activating reagentsuch as thionyl chloride, phosphorus oxychloride, EDC or EEDQ or thelike to generate the activated acyl compound 5.

In Scheme 8 Step 5, the poly-hydroxyl compound, compound 6, where Rrepresents an aromatic or an aliphatic cyclic or acyclic core, istreated with the activated acyl compound 5, in an appropriate solvent,optionally in the presence of a catalyst. Suitable catalysts includepyridine, dimethylaminopyridine, trimethylamine and the like to generatecompound 5. The catalyst can be used in quantities ranging from 0.01 to1.1 equivalents, relative to compound 3. Suitable solvents includemethylene chloride, ethyl acetate, diethyl ether, tetrahydrofuran,1,4-dioxane, 1,2-dimethoxyethane, toluene, combinations thereof and thelike. Reaction temperatures range from −10° C. to the boiling point ofthe solvent used; reaction completion times range from 1 to 96 h. Theactivated acyl compound 5 can be used in quantities ranging from 0.5 to15 equivalents relative to compound 6. The product, compound 7, can bepurified by methods known in the art.

The following examples are meant to illustrate the invention. They arenot meant to limit the invention in any way.

EXAMPLES Example 1. Preparation of Exemplary Acylated Active Agents

Compound 1:[(2R,3R)-5,7-di(butanoyloxy)-2-[3,4,5-tri(butanoyloxy)phenyl]chroman-3-yl]3,4,5-tri(butanoyloxy)benzoate

Butyryl chloride (6.03 mL) was added to a stirring solution of epigallocatechin gallate (2.0 g) and pyridine (6.28 mL) in dichloromethane (20mL) over 2 h between −5° C. to 5° C. The resulting mixture was stirredovernight at room temperature. The reaction mixture was then dilutedwith dichloromethane (100 mL), washed sequentially with water (50 mL),2N HCl (50 mL), saturated sodium bicarbonate (50 mL), and brine. Theorganic layer was evaporated in vacuo, and the resulting crude materialwas purified by flash chromatography by 30% ethyl acetate/heptane togive compound 1 (800 mg, 18%). ¹H NMR (CDCl₃): 7.6 (s, 2H), 7.22 (s,2H), 6.78 (s, 1H), 6.6 (s, 1H), 5.62 (t, 1H), 5.18 (s, 1H), 2.98-3.02(m, 2H), 2.4-2.6 (m, 16H), 1.6-1.8 (m, 16H), 0.92-1.02 (m, 24H).

Compound 2:[(2R,3R)-5,7-diacetoxy-2-(3,4,5-triacetoxyphenyl)chroman-3-yl]3,4,5-triacetoxybenzoate

Acetic anhydride (6.1 mL) was added dropwise to epigallo catechingallate (2.0 g) in pyridine (20 mL) at 0° C., and the resulting mixturewas stirred overnight at room temperature. Water was added to thereaction mixture, and the solid was filtered and washed with aq. 1N HCl(10 mL) and heptane (20 mL). The solid was then dissolved indichloromethane and passed through a silica gel filter column withdichloromethane as a mobile phase to furnish compound 2 (1.0 g, 28%)upon evaporation of volatiles. ¹H NMR (CDCl₃): 7.6 (s, 2H), 7.2 (s, 2H),6.75 (s, 1H), 6.6 (s, 1H), 5.6 (t, 1H), 5.19 (s, 1H), 2.98-3.02 (m, 2H),2.18-2.28 (m, 24H).

Compound 3:[(2R,3R)-5,7-bis(4-phenylbutanoyloxy)-2-[3,4,5-tris(4-phenylbutanoyloxy)phenyl]chroman-3-yl]3,4,5-tris(4-phenylbutanoyloxy)benzoate

Step 1:

To a solution of 4-phenylbutanoic acid (3 g, 18.27 mmol, 1 equiv.) andSOCl₂ (10.87 g, 91.35 mmol, 6.63 mL, 5 equiv.) in dichloromethane (50mL) is added one drop of DMF, then the mixture stirred at 20° C. for 5h. The solvent is removed in vacuum and toluene (20 mL) added to themixture. The mixture is concentrated in vacuo to afford 4-phenylbutanoylchloride (3.5 g, crude).

Step 2:

To a solution of[(2R,3R)-5,7-dihydroxy-2-(3,4,5-trihydroxyphenyl)chroman-3-yl]3,4,5-trihydroxybenzoate (1 g, 2.18 mmol, 1 equiv.) and K₂CO₃ (4.52 g,32.72 mmol, 15 equiv.) in acetonitrile (100 mL) was added a solution of4-phenylbutanoyl chloride (7.97 g, 43.63 mmol, 20 equiv.) inacetonitrile (10 mL), then the mixture was stirred at 20° C. for 10 h.The mixture was filtered, and the filtrate was concentrated in vacuum.The crude product was purified by silica gel chromatography (petroleumether/ethyl acetate=20:1-1:1) to afford compound 3 (2.2 g, 1.28 mmol,58.73% yield, 94.8% purity) as a white solid. LC/MS (M+H₃O⁺):1645.1

Compound 4: [2-acetoxy-4-(3,5,7-triacetoxy-4-oxo-chromen-2-yl)phenyl]acetate

To a mixture of 2-(3,4-dihydroxyphenyl)-3,5,7-trihydroxy-chromen-4-one(1 g) and acetic anhydride (2.36 g) in THF (40 mL) was added K₂CO₃ (3.2g) at 25° C., then the mixture was stirred at 55° C. for 12 h.Additional acetic anhydride was added (3 equiv.) and the mixture andstirred for another 3 h. The reaction mixture was concentrated in vacuumand purified by reverse phase prep-HPLC (C18; water (0.05% HCl)-ACNgradient) to give compound 4 (0.837 g, 49%) as a white solid. LCMS:513.2 (M+H⁺) ¹H NMR (400 MHz, CDCl₃). 7.742-7.703 (m, 2H), 7.373-7.346(m, 2H), 6.888 (s, 1H), 2.443, (s, 3H), 2.356 (s, 6H), 2.350 (s, 6H).

Compound 5:[2-butanoyloxy-4-[3,5,7-tri(butanoyloxy)-4-oxo-chromen-2-yl]phenyl]butanoate

To a mixture of 2-(3,4-dihydroxyphenyl)-3,5,7-trihydroxy-chromen-4-one(1 g) and butanoyl chloride (3.53 g) in THF (40 mL) was added TEA (3.35g) at 25° C., then the mixture was stirred at 55° C. for 12 h. Thereaction mixture was concentrated in vacuum and purified by reversephase prep-HPLC (C18, water (0.05% HCl)-ACN gradient) to give compound 5(1.13 g, 52% yield) as a colorless solid. LCMS: 653.3 (M+H⁺) ¹H NMR (400MHz, CDCl₃). 7.666-7.608 (m, 2H), 7.292-7.210 (m, 2H), 6.880 (s, 1H),2.542 (t, 2H), 2.535-2.484 (m, 8H), 1.753 (m, 10H), 1.020-0.997 (m,12H), 0.949 (t, 3H).

Compound 6: [2-octanoyloxy-4-[3,5,7-tri(octanoyloxy)-4-oxo-chromen-2-yl]phenyl] octanoate

To a mixture of 2-(3,4-dihydroxyphenyl)-3,5,7-trihydroxy-chromen-4-one(0.32 g) and octanoyl chloride (1.72 g) in THF (20 mL) was added TEA(1.07 g) at 25° C. Then the mixture was stirred at 55° C. for 12 h. Aportion of the solvent was removed in vacuum and the precipitate wascollected by filtration to give compound 6 (0.20 g, 20%) as a whitesolid. ¹H NMR (400 MHz, CDCl₃). 7.709-7.655 (m, 2H), 7.329-7.301 (m,2H), 6.837 (s, 1H), 2.723 (t, 2H), 2.612-2.539 (m, 8H), 1.751 (m, 10H),1.412-1.309 (m, 40H), 0.896 (m, 15H).

Compound 7:[2-decanoyloxy-4-[3,5,7-tris(decanoyloxy)-4-oxo-chromen-2-yl] phenyl]decanoate

To a mixture of 2-(3,4-dihydroxyphenyl)-3,5,7-trihydroxy-chromen-4-one(1 g) and decanoyl chloride (6.31 g) in THF (50 mL) was added TEA (3.35g) at 25° C., then the mixture was stirred at 55° C. for 12 h. A portionof the solvent was removed in vacuum and the precipitate was collectedby filtration to give compound 7 (2.47 g, 69%) as a white solid. ¹H NMR(400 MHz, CDCl₃). 7.772-7.669 (m, 2H), 7.343-7.321 (m, 2H), 6.685 (s,1H), 2.736 (t, 2H), 2.610-2.551 (m, 8H), 1.762 (m, 10H), 1.557-1.295 (m,50H), 0.899 (m, 15H).

Compound 8: [4-(3,5,7-triacetoxy-4-oxo-chromen-2-yl)phenyl] acetate

To a mixture of 3,5,7-trihydroxy-2-(4-hydroxyphenyl)chromen-4-one (2 g)in pyridine (15 mL) was added acetyl acetate (30 g), and then themixture was stirred at 15° C. for 12 hr under N2 atmosphere. The solventwas removed under reduced pressure and the residue was poured intocrushed ice with vigorous stirring. The solid precipitate was collectedby filtration and washed with cold water and then with methanol.Compound 8 (2.1 g, 65% yield) was obtained as a white solid. LCMS: 455.0(M+H⁺) ¹H NMR (400 MHz, CDCl₃) 7.858 (d, 2H), 7.339 (d, 1H), 7.278-7.257(m, 2H), 6.883 (d, 1H), 2.447 (s, 3H), 2.357 (s, 6H), 2.333 (s, 3H)

Compound 9: [4-(5,7-diacetoxy-4-oxo-chroman-2-yl)phenyl] acetate

5,7-dihydroxy-2-(4-hydroxyphenyl)chroman-4-one (0.500 g) was dissolvedwith pyridine (10 mL), and then acetyl acetate (0.844 g) was added tothe mixture reaction. The reaction mixture was stirred at 15° C. for 12h. The mixture reaction was concentrated under reduced pressure. Theresidue was purified by column chromatography (SiO₂, petroleumether/ethyl acetate gradient) to give compound 9 (0.300 g, 39% yield) asa white solid. LCMS: 416.1 (M+H₃O⁺) ¹H NMR (400 MHz, CDCl₃) 7.468 (d,2H), 7.166 (d, 2H), 6.793 (d, 1H), 6.551 (d, 1H), 5.497 (dd, 1H), 3.039(dd, 1H), 2.783 (dd, 1H), 2.393 (s, 3H), 2.326 (s, 3H), 2.308 (S, 3H).

Compound 10: [4-[5,7-di(butanoyloxy)-4-oxo-chroman-2-yl]phenyl]butanoate

To a solution of 5,7-dihydroxy-2-(4-hydroxyphenyl)chroman-4-one (0.500g) in pyridine (10 mL), was added butanoyl butanoate (1.02 g). Thereaction mixture was stirred at 15° C. for 12 h. The mixture wasconcentrated. The residue was purified by column chromatography (SiO₂,petroleum ether/ethyl acetate gradient) to give compound 10 (0.325 g,34% yield) as a white solid. LCMS: 500.2 (M+H30⁺) ¹H NMR (400 MHz,CDCl₃) 7.463 (d, 2H), 7.158 (d, 2H), 6.786 (d, 1H), 6.536 (d, 1H), 5.483(m, 1H), 3.031 (m, 1H), 2.662 (m, 1H), 2.586-2.524 (m, 6H), 1.837-1.785(m, 6H), 1.089-1.021 (m, 9H)

Compound 11:[3,5-diacetoxy-4-oxo-2-(3,4,5-triacetoxyphenyl)chromen-7-yl] acetate

To a solution of3,5,7-trihydroxy-2-(3,4,5-trihydroxyphenyl)chromen-4-one (1 g) inpyridine (10 mL) was added acetyl acetate (15.26 g), then the mixturewas stirred at 15° C. for 16 h. The solvent was removed and the mixturewas poured into ice water under stirring. The solid was filtered, washedwith water and dried in vacuum to give compound 11 (1.1 g, 61% yield) asa gray solid. LCMS 571.1 (M+H⁺) ¹H NMR (400 MHz, CDCl₃) 7.260 (s, 2H),7.349 (d, 1H), 6.886 (d, 1H), 2.441 (s, 3H), 2.372 (s, 3H), 2.353 (s,3H), 2.341 (s, 3H), 2.333 (s, 6H)

Compound 12:[4-[4-oxo-5,7-di(propanoyloxy)chromen-2-yl]-2-propanoyloxy-phenyl]propanoate

Propionic anhydride (1.33 mL, 10.4 mmol) was added dropwise to a stirredsolution of luteolin (0.3 g, 1.04 mmol) in anhydrous pyridine (2.5 mL,31.2 mmol) at 0° C. under N2 atmosphere. The resulting stirred solutionwas allowed to come to room temperature and reaction was monitored tocompletion by LCMS. The solution was diluted with 30 mL ethyl acetateand washed with H2O (30 mL), 1M HCl (30 mL), H2O (30 mL), and saturatedNaHCO3 (30 mL). The organic layer was dried over sodium sulfate,filtered, and concentrated by rotary evaporation. The crude residue waspurified by flash chromatography (silica, 10-100% ethyl acetate inhexanes) and fractions were concentrated by rotary evaporation to yieldcompounds (0.073 g, 15% yield) as an off-white solid. 1H-NMR (DMSO-d6,400 MHz): δ 12.75 (s, 1H), 8.07 (m, 2H), 7.5 (m, 1H), 7.15 (s, 1H), 7.12(d, 1H), 6.66 (d, 1H), 2.59-2.66 (m, 6H), 1.11-1.17 (m, 9H)

Compound 13:[4-oxo-3,5-di(propanoyloxy)-2-[3,4,5-tri(propanoyloxy)phenyl]chromen-7-yl]propanoate

Propionic anhydride (2 mL, 15.6 mmol) was added dropwise to a stirredsolution of myricetin (0.5 g, 1.56 mmol) in anhydrous pyridine (2.78 mL,47.1 mmol) at 0° C. under N₂ atmosphere. The resulting stirred solutionwas allowed to come to room temperature and reaction was monitored tocompletion by LCMS. The solution was diluted with 30 mL ethyl acetateand washed with H₂O (30 mL), 1 M HCl (30 mL), H₂O (30 mL), and saturatedNaHCO₃ (30 mL). The organic layer was dried over sodium sulfate,filtered, and concentrated by rotary evaporation. The crude residue waspurified by flash chromatography (silica, 10-100% ethyl acetate inhexanes) and fractions were concentrated by rotary evaporation to yieldCompound 13 (0.31 g, 30% yield) as a white solid. ¹H-NMR (DMSO-de, 400MHz): δ 7.77 (s, 2H), 7.64 (d, 1H), 7.16 (d, 1H), 2.60-2.70 (m, 12H),1.07-1.17 (m, 18H)

Compound 14:[4-[4-oxo-3,5,7-tri(propanoyloxy)chromen-2-yl]-2-propanoyloxy-phenyl]propanoate

Propionic anhydride (2.1 mL, 16.5 mmol) was added dropwise to a stirredsolution of quercetin (0.5 g, 1.65 mmol) in anhydrous pyridine (3.98 mL,49.5 mmol) at 0° C. under N2 atmosphere. The resulting stirred solutionwas allowed to come to room temperature and reaction was monitored tocompletion by LCMS. The solution was diluted with 30 mL ethyl acetateand washed with H2O (30 mL), 1M HCl (30 mL), H2O (30 mL), and saturatedNaHCO3 (30 mL). The organic layer was dried over sodium sulfate,filtered, and concentrated by rotary evaporation. The crude residue waspurified by flash chromatography (silica, 10-100% ethyl acetate inhexanes) and fractions were concentrated by rotary evaporation to yieldCompound 14 (0.1 g, 10% yield) as a white solid. 1H-NMR (DMSO-d6, 400MHz): δ 7.85 (m, 2H), 7.66 (d, 1H), 7.54 (d, 1H), 7.18 (d, 1H),2.62-2.89 (m, 10H), 1.09-1.19 (m, 20H)

Compound 15:[(2R,3R)-5,7-di(propanoyloxy)-2-[3,4,5-tri(propanoyloxy)phenyl]chroman-3-yl]3,4,5-tri(propanoyloxy)benzoate

Propionic anhydride (2.78 mL, 21.8 mmol) was added dropwise to a stirredsolution of epigallocatechin gallate (0.5 g, 1.09 mmol) in anhydrouspyridine (2.61 mL, 32.6 mmol) at 0° C. under N2 atmosphere. Theresulting stirred solution was allowed to come to room temperature andreaction was monitored to completion by LCMS. The solution was dilutedwith 30 mL ethyl acetate and washed with H2O (30 mL), 1M HCl (30 mL),H2O (30 mL), and saturated NaHCO3 (30 mL). The organic layer was driedover sodium sulfate, filtered, and concentrated by rotary evaporation.The crude residue was purified by flash chromatography (silica, 10-100%ethyl acetate in hexanes) and fractions were concentrated by rotaryevaporation to yield Compound 15 (0.695 g, 70% yield) as a white solid.1H-NMR (DMSO-d6, 400 MHz): δ 7.54 (s, 2H), 7.38 (s, 2H), 6.79 (m, 1H),6.66 (m, 1H), 5.66 (m, 1H), 5.54 (s, 1H), 3.13-3.17 (m, 1H), 2.96 (d,1H), 2.5-2.65 (m, 16H), 1.0-1.2 (m, 24H)

Compound 16: 5-amino-2-butanoyloxy-benzoic acid

Step 1:

To a mixture of 5-amino-2-hydroxy-benzoic acid (3 g, 19.59 mmol, 1equiv.) in methanol (50 mL) was added Boc₂O (4.28 g, 19.59 mmol, 4.50mL, 1 equiv.) in one portion at 15° C. under N2. The mixture was stirredat 15° C. for 5 h. The residue was poured into water (100 mL). Theaqueous phase was extracted with EtOAc (100 mL), and the organic phasewas dried with anhydrous Na₂SO₄, filtered, and concentrated in vacuo.The residue was used in next step without further purification.5-(tert-butoxycarbonylamino)-2-hydroxy-benzoic acid (4 g, crude) ascrude was obtained.

Step 2:

To a solution of 5-(tert-butoxycarbonylamino)-2-hydroxy-benzoic acid (4g, 15.79 mmol, 1 equiv.) and triethylamine (119.87 mg, 1.18 mmol, 164.88μL, 1 equiv.) in THF (30 mL) was added butanoyl chloride (126.22 mg,1.18 mmol, 123.74 μL, 1 equiv.) drop-wise at 0° C., while thetemperature was maintained below 0° C. The reaction mixture was warmedto 15° C. and stirred for 2 h. The reaction was quenched by slowaddition of ice, and then the mixture was extracted with EtOAc (100 mL).The organic phase was dried over anhydrous Na₂SO₄, filtered, andconcentrated in vacuo. The residue was purified by recrystallizationwith EtOAc (20 mL) to give the pure2-butanoyloxy-5-(tert-butoxycarbonylamino)benzoic acid (1.5 g, 4.64mmol, 29.37% yield) as white solid.

Step 3:

A solution of 2-butanoyloxy-5-(tert-butoxycarbonylamino)benzoic acid(1.5 g, 4.64 mmol, 1 equiv.) in HCl-EtOAc (20 mL, 4 M) was stirred at15° C. for 1 h. The mixture was filtered to obtain the product5-amino-2-butanoyloxy-benzoic acid as off-white solid (0.74 g, 2.76mmol, 59.58% yield, 96.993% purity, HCl salt). LC/MS: (M+H⁺) 224.1

Compound 17:2-butanoyloxy-5-[(E)-(4-butanoyloxy-3-carboxy-phenyl)azo]benzoic acid

A solution of[2-carboxy-4-[(E)-(3-carboxy-4-sodiooxy-phenyl)azo]phenoxy] sodium (2 g,5.78 mmol, 1 equiv.), butanoyl chloride (2.46 g, 23.11 mmol, 2.41 mL, 4equiv.), and NaOH (462.12 mg, 11.55 mmol, 2 equiv.) in DMF (100 mL) wasstirred at 50° C. for 0.5 h. The solid was filtered, water (150 mL) wasadded to the filtrate, and the mixture was filtered again. The resultingsolids filter cake was dried in vacuo.2-butanoyloxy-5-[(E)-(4-butanoyloxy-3-carboxy-phenyl)azo]benzoic acid(0.8 g) was obtained as brown solid. LC/MS: (M+H⁺): 443.1

Compound 18:5-amino-2-[(2R,3R,4S,5R)-3,4,5-tri(butanoyloxy)tetrahydropyran-2-yl]oxy-benzoicacid

Xylose tributyrate (1.87 g, 5.18 mmol, 1 eq), tert-butyl5-((tert-butoxycarbonyl)amino)-2-hydroxybenzoate (2.4 g, 7.8 mmol, 1.5eq) and triphenylphosphine (2.05 g, 7.8 mmol, 1.5 eq) were dissolved inanhydrous THF (37.5 mL) under N2 and cooled to 0° C. Di-tert-butylazodicarboxylate (1.8 g, 7.8 mmol, 1.5 eq) was added and the reactionwas stirred at 0° C. for 1 hour under N2. The ice bath was removed andthe reaction was stirred at room temperature overnight. The reactionmixture was loaded directly onto silica and dried by rotary evaporation.The solid loaded sample was purified by multiple rounds of columnchromatography (gradient: 0-50% ethyl acetate in hexanes) to separatethe anomers and yield(2S,3R,4S,5R)-2-(2-(tert-butoxycarbonyl)-4-((tert-butoxycarbonyl)amino)phenoxy)tetrahydro-2H-pyran-3,4,5-triyltributyrate (45 mg). The purified compound was dissolved in chloroform(1 mL), followed by addition of 4 M HCl in dioxane (1.2 mL). Afterdeprotection was complete as confirmed by LCMS, the solution wasconcentrated by rotary evaporation and dried under high vacuum overnightto yield the title compound (11 mg, 0.021 mmol, 0.4% yield). LCMS[M−H]⁻: 494.5. ¹H NMR (400 MHz, DMSO-d₆) δ 6.83-6.75 (m, 2H), 6.64-6.53(m, 1H), 5.58 (d, J=3.6 Hz, 1H), 5.53 (t, J=9.9 Hz, 1H), 5.03-4.90 (m,2H), 3.89 (t, J=10.9 Hz, 1H), 3.73 (dd, J=10.9, 5.9 Hz, 1H), 2.38-2.12(m, 6H), 1.58-1.39 (m, 6H), 0.92-0.76 (m, 9H).

Compound 19:5-amino-2-[(2S,3R,4S,5R)-3,4,5-tri(butanoyloxy)tetrahydropyran-2-yl]oxy-benzoicacid

Step 1

5-Amino salicylic acid (10.0 g) was dissolved in a mixture of dioxane(100 mL), water (100 mL), and NaOH (2.60 g), and the resulting solutionwas cooled in an ice-bath. Di-tert-butyl dicarbonate (Boc anhydride)(15.60 g) was added, and the mixture was warmed to room temperature andstirred for 1.0 h. The solution was concentrated to 60 mL, diluted withethyl acetate (100 mL), and the resulting mixture was cooled in anice-bath. The mixture was acidified with aq. KHSO₄ to pH 2-3. Theaqueous layer was extracted with EtOAc. The organic phase was washedwith water, brine, dried over Na₂SO₄, filtered, and concentrated toafford 5-(tert-butoxycarbonylamino)-2-hydroxy-benzoic acid (7.0 g, 42%).

Step 2

5-(tert-butoxycarbonylamino)-2-hydroxy-benzoic acid (3 g) was dissolvedin DMF, and the resulting solution was cooled to 0° C.1,1′-Carbonyldiimidazole (CDI) was added, and the mixture was stirred atroom temperature for 2 h. Then, tert-butylalcohol (1.7 g) and DBU (2.1g) were added. The reaction was stirred at room temperature overnight.The reaction mixture was poured onto ice-water, and the solid product,tert-butyl 5-(tert-butoxycarbonylamino)-2-hydroxy-benzoate, wascollected by filtration (3.0 g, 81.9%).

Step 3

To a mixture of tert-butyl5-(tert-butoxycarbonylamino)-2-hydroxy-benzoate,[(3R,4S,5R)-4,5-di(butanoyloxy)-6-hydroxy-tetrahydropyran-3-yl]butanoate (1.2 g) and triphenylphosphene (1.2 g) in THF (50 mL) wasadded di-t-butyl azodicarboxylate (DTAD) (1.1 g) DTAD, and the mixturewas stirred overnight at room temperature. The product was purified byreverse phase chromatography using acetonitrile-water to affordtert-butyl5-(tert-butoxycarbonylamino)-2-[(3R,4S,5R)-3,4,5-tri(butanoyloxy)tetrahydropyran-2-yl]oxy-benzoateas sticky solid (0.6 g, 30.0%).

Step 4

Tert-butyl5-(tert-butoxycarbonylamino)-2-[(3R,4S,5R)-3,4,5-tri(butanoyloxy)tetrahydropyran-2-yl]oxy-benzoate(600 mg) was added to 4M HCl in dioxane (15 mL) and stirred at roomtemperature overnight. After the consumption of the starting material,the organic phase was evaporated, and the residue was co-evaporated withheptane and dichloromethane twice more. The solid obtained was driedunder high vacuum to afford compound the title product as dark brownsolid (200 mg, 43.8%). Fractionation of the product afforded twoanomeric isomers (Compounds 21 and 22). ¹H NMR (DMSO d6): Isomer 1: δ7.62 (d, 1H), 7.45 (dd, 1H), 7.38 (d, 1H), 6 (d, 1H), 5.6 (t, 1H),5.0-5.1 (m, 1H), 4.7-4.75 (m, 1H), 3.6-3.8 (m, 1H), 3.45-3.6 (1H),2.1-2.3 (m, 6H), 1.4-1.6 (m, 6H), 0.75-0.85 (m, 9H). Isomer 2: δ 7.82(d, 1H), 7.5 (dd, 1H), 7.05 (d, 1H), 5.5 (d, 1H), 5.3 (t, 1H), 5.1-5.15(m, 1H), 4.9-5.0 (m, 1H), 4.0-4.08 (m, 1H), 3.7-3.8 (1H), 2.1-2.3 (m,6H), 1.4-1.6 (m, 6H), 0.75-0.85 (m, 9H) ppm

Compound 20: 5-(butanoylamino)-2-hydroxy-benzoic acid

To the solution of 5-amino-2-hydroxy-benzoic acid (1 g) andtriethylamine (0.991 g) in dioxane (20 mL) and H₂O (10 mL) was addedbutyric anhydride (1.24 g), and the mixture was stirred at 20° C. for 16h. The dioxane was removed under reduced pressure and the pH wasadjusted to 5-6 by adding aqueous 3N HCl at 0° C. The solid wasfiltered, washed three times with water (20 mL) and concentrated invacuum. The crude product was purified by reverse phase prep-HPLC (C18,water (0.05% HCl)-acetonitrile gradient) to give5-(butanoylamino)-2-hydroxy-benzoic acid (0.3 g, 20%) as a light pinksolid. LCMS: 224.1 (M+H⁺) ¹H NMR (400 MHz, DMSO-d6): δ 11.015 (br, 1H),9.807 (s, 1H), 8.106 (d, 1H), 7.652 (dd, 1H), 6.893 (d, 1H), 2.239 (m,2H), 1.604 (m, 2H), 0.902 (t, 3H) ppm

Compound 21:5-amino-2-(((2R,3R,4R,5R)-3,4,5-tris(butyryloxy)tetrahydro-2H-pyran-2-yl)oxy)benzoicacid Step 1. Ribose Tetrabutyrate

To a stirred solution of D-(+)-ribose 1 (5 g) in anhydrous pyridine(24.2 mL) was added solution of butyryl chloride (23.70 g) indichloromethane (50 mL) at 0-5° C. The reaction mixture was brought toroom temperature and stirred for 16 h. The mixture was diluted withdichloromethane (100 ml) and washed successively with water (100 mL), 2Naqueous HCl (300 mL), saturated sodium bicarbonate solution (300 mL) andbrine (100 mL). The organic layer was dried over sodium sulphate andconcentrated under reduced pressure. The residue was purified by columnchromatography over silica gel (5-10% EtOAc-hexane gradient) to affordribose tetrabutyrate as a colorless oil (7.5 g, 52%, mixture of α/βanomers).

Step 2. Ribose Tributyrate

Ammonium hydroxide (11 mL) was added slowly to a mixture of ribosetetrabutyrate 2 (7.5 g) in acetonitrile (60 mL) at room temperature andthe resulting reaction mixture was stirred for 5 h. The mixture wasdiluted with MTBE (75 mL) and stirred for 15 minutes. The organic layerwas separated and concentrated under reduced pressure and the residuewas partitioned between MTBE (100 mL) and water (75 mL). The MTBE layerwas separated, dried over sodium sulphate and concentrated under reducedpressure. The residue was purified by column chromatography [usingsilica gel 100-200 mesh and 10-20% EtOAc-Hexane as eluting solvent] toafford ribose tributyrate as a colorless oil (1.1 g, 17%).

Step 3. 5-tert-butoxycarbonylamino-2-hydroxy-benzoic acid

To the stirred solution of 5-amino salicylic acid 4 (5 g) in 1,4-dioxaneand water (1:1; 100 mL) was added NaOH (1.3 g) and Boc-anhydride (7.83g) at 0° C. and the resulting reaction mixture was stirred at roomtemperature for 1 h. The reaction mixture was concentrated under reducedpressure, the residue was diluted with EtOAc (50 mL) and the pH wasadjusted to 3-4 by dropwise addition of 0.5N aqueous HCl at 0° C. Theorganic layer was separated and aqueous layer was extracted with EtOAc(50 mL). The combined organic layer was dried over sodium sulphate andconcentrated under reduced pressure to provide5-tert-butoxycarbonylamino-2-hydroxy-benzoic acid as off white solid(5.3 g, 64%).

Step 4. 5-tert-butoxycarbonylmethyl-2-hydroxy-benzoic acid tert-butylester

To a stirred solution of 5-tert-butoxycarbonylamino-2-hydroxy-benzoicacid 5 (5.3 g) in DMF (50 mL) was added CDI (3.39 g) at 0-5° C. andstirred for 2 h. tert-Butanol (4.025 mL) and DBU (2.54 mL) were thenadded and the mixture was stirred at room temperature for 16 h. Themixture was diluted with water (100 mL) and extracted with EtOAc (200mL). The organic layer was separated, dried over sodium sulphate andconcentrated under reduced pressure. The residue was purified by columnchromatography using silica gel [100-200 mesh; under gradient elution of5-10% EtOAc-Hexane] to afford5-tert-butoxycarbonylmethyl-2-hydroxy-benzoic acid tert-butyl ester asoff white solid (2 g, 31%).

Step 5.5-tert-butoxycarbonylamino-2-(3,4,5-tris-butyryloxy-tetrahydro-pyran-2-yloxy)-benzoicacid tert-butyl ester

To a stirred solution of 5-tert-butoxycarbonylmethyl-2-hydroxy-benzoicacid tert-butyl ester 6 (0.850 g) and ribose tributyrate (1.04 g) in THF(5 mL) was sequentially added triphenylphosphine (1.03 g) anddi-tert-butyl azodicarboxylate (0.948 g) at room temperature and themixture was stirred for 16 h. The mixture was concentrated under reducedpressure and the residue was purified by column chromatography oversilica gel (5 to 18% EtOAc-Hexane gradient) to afford of crude5-tert-butoxycarbonylamino-2-(3,4,5-tris-butyryloxy-tetrahydro-pyran-2-yloxy)-benzoicacid tert-butyl ester (1.3 g) which was used directly in the next step.

Step 6.5-amino-2-[(3R,4R,5R)-3,4,5-tri(butanoyloxy)tetrahydropyran-2-yl]oxy-benzoicacid

To a stirred solution of crude5-tert-butoxycarbonylamino-2-(3,4,5-tris-butyryloxy-tetrahydro-pyran-2-yloxy)-benzoicacid tert-butyl ester 7 (1.3 g, crude from above experiment) in1,4-dioxane (7 mL) was added 4N HCl in 1,4-dioxane (10 mL) at 0° C. andthe resulting reaction mixture was stirred at room temperature for 16 h.Then reaction mixture was concentrated under reduced pressure and theresidue was purified by reverse phase prep-HPLC to provide5-amino-2-(((2R,3R,4R,5R)-3,4,5-tris(butyryloxy)tetrahydro-2H-pyran-2-yl)oxy)benzoicacid (0.05 g). LCMS: 496.5 (M+H⁺) ¹H NMR (400 MHz, DMSO-d6): δ6.919-6.898 (m, 2H), 6.658 (m, 1H), 5.431 (m, 1H), 5.350 (m, 1H), 5.234(m, 1H), 5.161 (m, 1H), 4.213 (m 1H), 3.749 (m, 1H), 2.497-2.268 (m,4H), 2.197 (m, 1H), 1.620-1.487 (m, 6H), 0.926-0.888 (m, 9H) ppm

Compound 22:5-amino-2-[(2R,3R,4S,5S)-3,4,5-tri(butanoyloxy)tetrahydropyran-2-yl]oxy-benzoicacid

Step 1: 2-Hydroxy-4-nitro-benzoate (20 g) and KHCO₃ (13.1 g) weresuspended in DMF (100 mL). To the suspension was added benzyl bromide(22.4 g) and the reaction mixture was stirred at room temperatureovernight. Water (150 mL) was added and the resulting mixture wasextracted with ethyl acetate (250 mL). The organic phase was separatedand washed twice with water, brine, and dried over Na₂SO₄. The solventwas removed under reduced pressure and the residue was purified bycolumn chromatography (hexanes/ethyl acetate gradient).Recrystallization from 15% ethyl acetate in hexanes provided benzyl2-hydroxy-4-nitro-benzoate (10.5 g).

Step 2: Benzyl 2-hydroxy-4-nitro-benzoate (8.5 g), arabinose tributyrate(7.5 g) and triphenylphosphine (8.2 g) were dissolved in THF (150 mL)and stirred at 0° C. To this mixture was added di-t-butylazodicarboxylate (7.2 g) and stirring was continued at 0° C. for 1 h,then at room temperature overnight. The reaction mixture wasconcentrated and purification by column chromatography (hexanes/ethylacetate gradient) provided benzyl5-nitro-2-[(2R,3R,4S,5S)-3,4,5-tri(butanoyloxy)tetrahydropyran-2-yl]oxy-benzoate(1.78 g, 14%).

Step 3:5-nitro-2-[(2R,3R,4S,5S)-3,4,5-tri(butanoyloxy)tetrahydropyran-2-yl]oxy-benzoate(0.095 g) was dissolved in methanol (15 mL) and stirred at roomtemperature. To this mixture was added 10% Pd/C (0.05 g). The suspensionwas stirred under a hydrogen atmosphere at room temperature overnight.The reaction mixture was filtered through Celite and washed withmethanol. The combined filtrate and washing were concentrated. Theresidue was purified by reverse phase chromatography (C-18, 0.1%trifluoroacetic acid in acetonitrile and 0.1% trifluoroacetic acid inwater) to give5-amino-2-[(2R,3R,4S,5S)-3,4,5-tri(butanoyloxy)tetrahydropyran-2-yl]oxy-benzoicacid (0.045 g, 59%). MS 494.2 (M−H) NMR (DMSO d6): δ 7.223 (m, 1H),7.139 (m, 1H), 6.997 (s, 1H), 7.851 (d, 1H), 5.469 (m, 1H), 5.350 (m,1H), 5.239 (m, 1H) 4.127 (d, 1H), 3.672 (d, 1H), 2.490-2.369 (M, 6H),1.596-1.485 (m, 6H), 0.924-0.818 (m, 9H) ppm

Compound 23:2-(((2S,3S,4R,5R,6S)-3,4,5-tris(butyryloxy)-6-methyltetrahydro-2H-pyran-2-yl)oxy)benzoicacid

Step 1:

To a mixture of[(2S,3R,4R,5S)-4,5-di(butanoyloxy)-6-hydroxy-2-methyl-tetrahydropyran-3-yl]butanoate(0.95 g, 2.54 mmol) and tert-butyl 2-hydroxybenzoate (0.468 g, 2.41mmol) in THF (10 mL) was added tert-butyl(NE)-N-tert-butoxycarbonyliminocarbamate (0.876 g, 3.81 mmol) and PPh₃(0.952 g, 3.63 mmol) in one portion at 15° C. under N2. The mixture wasstirred at 15° C. for 12 h. The reaction mixture was filtered andconcentrated under reduced pressure to give a residue which was purifiedby prep-HPLC [water (10 mM NH₄HCO₃)-ACN] to give tert-butyl2-[(3S,4R,5R,6S)-3,4,5-tri(butanoyloxy)-6-methyl-tetrahydropyran-2-yl]oxybenzoate(0.3 g, 0.544 mmol, 21% yield) as a white solid.

Step 2:

To a solution of tert-butyl2-[(3S,4R,5R,6S)-3,4,5-tri(butanoyloxy)-6-methyl-tetrahydropyran-2-yl]oxybenzoate(0.15 g, 0.272 mmol) in DCM (5 mL) was added TFA (0.031 g, 0.27 mmol).The mixture was stirred at 15° C. for 12 hr. The reaction mixture wasfiltered and concentrated under reduced pressure to give a residue. Theresidue was purified by prep-HPLC [water (0.1% TFA)-ACN] to givecompound 23 and compound 29.

Compound 23 was prepared as a colorless oil (0.003 g, 1.9% yield). LCMS:517.2 (M+Na⁺); ¹H NMR CDCl3 8.192 (m, 1H), 7.565 (m, 1H0, 7.441 (m, 1H),7.255 (m, 1H), 5.813 (m, 1H), 5.525-5.444 (m, 3H), 4.413 (m, 1H), 2.460(t, 2H), 2.356 (t, 2H), 2.233 (t, 2H), 1.627 (m, 6H), 1.225 (d, 3H),1.028 (t, 3H), 0.938 (t, 3H), 0.919 (t, 3H)

Compound 24 was synthesized in a similar manner to compound 22.

Compound 24:2-(((2R,3R,4S,5R,6R)-3,4,5-tris(butyryloxy)-6-((butyryloxy)methyl)tetrahydro-2H-pyran-2-yl)oxy)benzoicacid

Compound 25:2-(((3S,4R,5R,6S)-6-methyl-3,4,5-tris(propionyloxy)tetrahydro-2H-pyran-2-yl)oxy)benzoicacid

Step 1

To a mixture of[(2S,3R,4R,5S)-6-hydroxy-2-methyl-4,5-di(propanoyloxy)tetrahydropyran-3-yl]propanoate(1 g, 3.01 mmol) and tert-butyl 2-hydroxybenzoate (1.17 g, 6.02 mmol) inTHF (10 mL) was added di-tert-butyl azodicarboxylate (1.04 g, 4.51 mmol)and triphenylphosphine (2.77 g, 4.51 mmol) in one portion at 15° C.under N₂. The mixture was stirred at 15° C. for 12 h. The reactionmixture was filtered and concentrated under reduced pressure and theresidue was purified by prep-HPLC. [water (10 mM NH₄HCO₃)-ACN] to givetert-butyl2-[(3S,4R,5R,6S)-6-methyl-3,4,5-tri(propanoyloxy)tetrahydropyran-2-yl]oxybenzoate(0.6 g, 39% yield) as a yellow solid.

Step 2

To a mixture of tert-butyl2-[(3S,4R,5R,6S)-6-methyl-3,4,5-tri(propanoyloxy)tetrahydropyran-2-yl]oxybenzoate(0.44 g, 0.865 mmol) in DCM (5 mL) was added TFA (0.099 g, 0.865 mmol)in one portion at 15° C. under N₂. The mixture was stirred 12 h. Thereaction mixture was filtered and concentrated under reduced pressure togive a residue. The residue was purified by prep-HPLC [water (0.1%TFA)-ACN].2-[(3S,4R,5R,6S)-6-methyl-3,4,5-tri(propanoyloxy)tetrahydropyran-2-yl]oxybenzoicacid (0.067 g, 15% yield) as a white solid. MS 475.1 (M+Na) NMR (DMSOd6): δ 8.1 (m, 1H), 7.5 (m, 1H), 7.1 (m, 2H), 5.6 (m, 1H), 5.4 (m, 1H),5.3 (m, 1H), 5.1 (m, 1H) 4.0 (m, 1H), 2.2 (m, 6H), 1.2 (m, 12H).

Compound 26:2-(((2S,3R,4S,5S)-3,4,5-tris(propionyloxy)tetrahydro-2H-pyran-2-yl)oxy)benzoicacid

Step 1

To a solution of (3R,4S,5S)-2-hydroxytetrahydro-2H-pyran-3,4,5-triyltripropionate (188.47 mg, 592.09 umol, 1 eg), tert-butyl2-hydroxybenzoate (230 mg, 1.18 mmol, 2 eg) and triphenylphosphine(545.97 mg, 888.14 umol, 1.5 eg) in THF (10 mL) was addeddi-tertbutyazodicarboxylate (204.50 mg, 888.14 umol, 1.5 eg) at 0° C.The mixture was stirred at 15° C. for 16 hr. TLC indicated new spotformed. The reaction mixture was concentrated under reduced pressure togive a residue. The residue was purified by column chromatography (SiO₂,Petroleum ether/Ethyl acetate=10:1 to 1:1). Compound tert-butyltert-butyl2-[(3R,4S,5S)-3,4,5-tri(propanoyloxy)tetrahydropyran-2-yl]oxybenzoate(200 mg, 404.42 umol, 68.30% yield) was obtained as a white solid.

Step 2

To a solution of tert-butyl2-[(3R,4S,5S)-3,4,5-tri(propanoyloxy)tetrahydropyran-2-yl] oxybenzoate(100 mg, 202.21 umol, 1 eg) in CH₂Cl₂ (1 mL) was added TFA (138.34 mg,1.21 mmol, 89.83 uL, 6 eg) at 15° C. The mixture was stirred at 15° C.for 2 hr. LCMS showed desired MS was detected. The reaction mixture wasconcentrated under reduced pressure to give a residue. The residue waspurified by prep-TLC (SiO₂, Petroleum ether/Ethyl acetate=1:1). Compound2-[(3R,4S,5S)-3,4,5-tri(propanoyloxy)tetrahydropyran-2-yl]oxybenzoicacid (30 mg, 32.84 μmol, 16.24% yield, 46.66% purity) was obtained as acolorless oil. LCMS: (M−1) 437.1 NMR (CDCl₃): 58.2 (m, 1H), 7.4 (m, 1H),7.2 (m, 2H), 5.3 (m, 4H), 4.0 (dd, 2H), 2.47-1.1 (m, 9H) ppm.

Compound 27:2-(((2S,3R,4S,5R)-3,4,5-tris(propionyloxy)tetrahydro-2H-pyran-2-yl)oxy)benzoicacid

Step 1:

To a solution of[(3R,4S,5R)-6-hydroxy-4,5-di(propanoyloxy)tetrahydropyran-3-yl]propanoate (0.500 g, 1.57 mmol), tert-butyl 2-hydroxybenzoate (0.610 g,3.14 mmol) and PPh₃ (0.824 g, 3.14 mmol) in THF (10 mL) was addeddi-tert-butyl azodicarboxylate (0.723 g, 3.14 mmol) at 0° C. Thereaction was stirred for 12 h at 15° C. The mixture reaction wasconcentrated under reduced pressure. The residue was purified by columnchromatography (SiO₂, petroleum ether/ethyl acetate=30/1 to 5:1) to givetert-butyl 2-[(3R, 4S,5R)-3,4,5-tri(propanoyloxy)tetrahydropyran-2-yl]oxybenzoate (0.320 g,37% yield) as a brown solid.

Step 2:

To a solution of TFA (10 mL) in CH₂Cl₂ (30 mL) was added tert-butyl2-[(3R,4S,5R)-3,4,5-tri(propanoyloxy)tetrahydropyran-2-yl]oxybenzoate(0.300 g, 606 mmol) and the mixture was stirred at 15° C. for 2 h. Themixture reaction was concentrated under reduced pressure. The residuewas purified by prep-HPLC (column: Waters Xbridge Prep OBD C18 150*40 10u; mobile phase: [water (10 mM NH4HCO3)-ACN]; B %: 1%-50%,11 min) togive2-[(3R,4S,5R)-3,4,5-tri(propanoyloxy)tetrahydropyran-2-yl]oxybenzoicacid (0.010 g, 3.4% yield) as a yellow oil. MS 437.1 (M−H) NMR (DMSOd6): δ 7.5 (m, 1H), 7.3 (m, 1H), 7.1 (m, 1H), 7.0 (m, 1H), 5.9 (m, 1H),5.6 (1H) 5.0 (m, 2H), 4.0 (m, 1H), 3.7 (m, 1H), 2.2 (m, 6H), 0.97 (m,9H).

Compound 28:2-(((2S,3R,4S,5R,6R)-3,4,5-tris(butyryloxy)-6-((butyryloxy)methyl)tetrahydro-2H-pyran-2-yl)oxy)benzoicacid

To the solution of 2-hydroxybenzoic acid (6 g, 43.44 mmol, 7.50 mL, 1eq) and CDI (8.45 g, 52.13 mmol, 1.2 eq) in DMF (50 mL) was added DBU(7.94 g, 52.13 mmol, 7.86 mL, 1.2 eq) and t-BuOH (6.47 g, 87.32 mmol,8.35 mL, 2.01 eq). The mixture was stirred at 15° C. for 16 h. LCMS(ET14826-364-P1A) showed the reaction was completed. The solvent wasremoved under reduced pressure. The crude product was purified by silicagel chromatography eluted with Petroleum ether/Ethyl acetate=1:0-2:1 togive tert-butyl 2-hydroxybenzoate (5 g, 25.74 mmol, 59.26% yield)

To the solution of (2R,3S,4R,5R)-2,3,4,5,6-pentahydroxyhexanal (20 g,111.02 mmol, 1 eq) in DCM (500 mL) was added butyryl chloride (94.63 g,888.12 mmol, 92.77 mL, 8 eq) and the mixture was stirred at 15° C. for0.5 h. Then pyridine (70.25 g, 888.12 mmol, 71.68 mL, 8 eq) was added tothe solution dropwise slowly. After the addition, the mixture wasstirred at 15° C. for another 16 h. LCMS (ET14826-367-P1A) showed thereaction was completed. The solvent was removed under reduced pressure.The crude product was purified by silica gel chromatography eluted withPetroleum ether/Ethyl acetate=1:0-5:1 to give(3R,4S,5R,6R)-6-((butyryloxy)methyl) tetrahydro-2H-pyran-2,3,4,5-tetrayltetrabutyrate (58 g, 109.31 mmol, 98.46% yield) as yellow oil

To the solution of(3R,4S,5R,6R)-6-((butyryloxy)methyl)tetrahydro-2H-pyran-2,3,4,5-tetrayltetrabutyrate (10 g, 18.85 mmol, 1 eq) in THF (85 mL) and H₂O (5 mL) wasadded methanamine/THF (2 M, 12.25 mL, 1.3 eg). Then the mixture wasstirred at 15° C. for 16 h. LCMS (ET14826-370-P1A2) showed most of thestarting material was consumed and the desired MS was detected. Thesolvent was removed under reduced pressure. The crude product waspurified by silica gel chromatography eluted with Petroleum ether/Ethylacetate=10:1-1:1 to give(2R,3R,4S,5R)-2-((butyryloxy)methyl)-6-hydroxytetrahydro-2H-pyran-3,4,5-triyltributyrate (10 g, 21.50 mmol, 57.03% yield, 99% purity) as yellow oil.

To the solution of(2R,3R,4S,5R)-2-((butyryloxy)methyl)-6-hydroxytetrahydro-2H-pyran-3,4,5-triyltributyrate (0.85 g, 1.85 mmol, 1 eg) and tert-butyl 2-hydroxybenzoate(340.57 mg, 1.75 mmol, 0.95 eg) in THF (20 mL) was added PPh₃ (692.29mg, 2.64 mmol, 1.43 eg) and di-tert-butyl azodicarboxylate (637.51 mg,2.77 mmol, 1.5 eg). Then the mixture was stirred at 15° C. for 16 h.LCMS showed the reaction was completed and the desired MS was detected.The solvent was removed under reduced pressure. The crude product waspurified by p-HPLC (column: Phenomenex Luna C18 200*40 mm*10 um; mobilephase: [water (0.1% TFA)-ACN]; B %: 75%-95%,10 min) to give(3R,4S,5R,6R)-2-(2-(tert-butoxycarbonyl)phenoxy)-6-((butyryloxy)methyl)tetrahydro-2H-pyran-3,4,5-triyltributyrate (0.2 g, 314.11 umol, 17.02% yield) as brown oil.

To the solution of(3R,4S,5R,6R)-2-(2-(tert-butoxycarbonyl)phenoxy)-6-((butyryloxy)methyl)tetrahydro-2H-pyran-3,4,5-triyl tributyrate (0.2 g, 314.11 umol, 1 eg)in DCM (10 mL) was added TFA (1.54 g, 13.51 mmol, 1 mL, 43.00 eg). Thenthe solution was stirred at 15° C. for 16 h. LCMS showed the reactionwas completed and the desired MS was detected. The solvent was removedunder reduced pressure. The crude product was purified by p-HPLC(column: Nano-micro Kromasil C18 100*30 mm um; mobile phase: [water(0.1% TFA)-ACN]; B %: 60%-78%,10 min) to afford the title compound (24mg, 40.10 umol, 12.76% yield, 97% purity, temporary assigned) as yellowoil. LCMS: (M+18): 598.2 NMR (DMSO d6): δ 8.1 (d, 1H), 7.5 (dd, 1H), 7.4(d, 1H), 7.2 (m, 1H), 5.8 (m, 1H), 5.6 (t, 1H), 5.2 (m, 2H), 4.1 (m,3H), 2.3 (m, 8H), 1.6 (m, 8H), 0.87 (m, 12H) ppm.

Compound 29:2-(((3S,4R,5R,6S)-3,4,5-tris(butyryloxy)-6-methyltetrahydro-2H-pyran-2-yl)oxy)benzoicacid

Compound 29 was prepared as a colorless oil (0.03 g, 22% yield). LCMS:517.2 (M+Na⁺); ¹H NMR CDCl3 8.080 (m, 1H), 7.491 (m, 1H), 7.164 (m, 1H),7.086 (m, 1H), 5.494 (m, 1H), 5.292 (m, 1H), 5.197 (m, 1H), 5.133 (m,1H), 3.946 (m, 1H), 2.413-2.153 (m, 6H), 1.653-1.546 (m, 6H), 1.204 (d,3H), 0.946 (t, 3H), 0.869 (t, 3H), 0.850 (t, 3H).

Compound 30:2-(((2S,3R,4S,5S)-3,4,5-tris(butyryloxy)tetrahydro-2H-pyran-2-yl)oxy)benzoicacid

Compound 30 was prepared in an analogous matter to compound 29. Compound30 was prepared in 27% yield (15 mg). LCMS: 503 (M+Na⁺); ¹H NMR CDCl38,095 (m, 1H), 7.484 (m, 1H), 7.326 (m, 1H), 7.154 (m, 1H), 5.775 (d,1H), 5.482-5.404 (m, 3H), 4.084 (d, 1H), 3.840 (d, 1H), 2.372-2.276 (m,6H), 1.658-1.497 (m, 6H), 0.937 (t, 3H), 0.863 (t, 3H), 0.823 (t, 3H).

Compound 31:2-(((2R,3R,4S,5S)-3,4,5-tris(butyryloxy)tetrahydro-2H-pyran-2-yl)oxy)benzoicacid

Step 1:

A solution of DCC (8 g, 38.8 mmol) in THF (50 mL) was added dropwise toa solution of 2-hydroxybenzoic acid (5 g, 36.2 mmol) and DMAP (0.17 g,1.39 mmol) in t-BuOH (100 mL) and the mixture was stirred at 15° C. for16 h. The crude product was purified by silica gel chromatography(petroleum ether/ethyl acetate=1:0) to give tert-butyl 2-hydroxybenzoate(3 g, 42.7%) as colorless oil.

Step 2:

To a solution of (3R,4S,5S)-2-hydroxytetrahydro-2H-pyran-3,4,5-triyltributyrate (0.7 g, 1.94 mmol) and tert-butyl 2-hydroxybenzoate (0.358g, 1.85 mmol) in THF (30 mL) was added PPh₃ (0.728 g, 2.78 mmol) andDBAD (0.671 g, 2.91 mmol) in portions. Then the mixture was stirred at15° C. for 16 h. The solvent was removed under reduced pressure. Thecrude product was purified by prep-HPLC (column: Agela innoval ods-2250*80 mm; mobile phase: [water (0.1% TFA)-ACN]; B %: 37%-67%,20 min) togive(3R,4S,5S)-2-(2-(tert-butoxycarbonyl)phenoxy)tetrahydro-2H-pyran-3,4,5-triyltributyrate (0.4 g, 0.745 mmol, 38.4%) as a yellow oil.

Step 3:

To a solution of(3R,4S,5S)-2-(2-(tert-butoxycarbonyl)phenoxy)tetrahydro-2H-pyran-3,4,5-triyltributyrate (0.2 g, 0.37 mmol) in DCM (10 mL) was added TFA (3.08 g, 27mmol). Then the mixture was stirred at 15° C. for 16 h under N2. Thesolvent was removed under reduced pressure. The crude product waspurified by prep-HPLC (column: Waters Xbridge 150*25 5 u; mobile phase:[water (0.1% TFA)-ACN]; B %: 47%-67%, 12 min) to give the titlecompound. LCMS: 503 (M+Na⁺); ¹H NMR CDCl3 8.202 (m, 1H), 7.567 (m, 1H),7.264-7.213 (m, 2H), 5.480-5.351 (m, 4H), 4.061 (m, 1H), 3.8 (m, 1H),2.425-2.297 (m, 6H), 1.688-1.650 (m, 6H), 0.959 (m, 9H)

Compound 32:5-amino-2-(((2S,3R,4S,5R)-3,4-bis(butyryloxy)-5-hydroxytetrahydro-2H-pyran-2-yl)oxy)benzoicacid

Pancreatin from porcine pancreas (200 mg) and FaSSIF/FeSSIF/FaSSGFpowder (44.8 mg, sourced from biorelevant.com) were suspended in 20 mLof SIF buffer (10.5 mM sodium hydroxide, 28.6 mM monobasic sodiumphosphate monohydrate, 106 mM sodium chloride, pH 6.5) and incubated at37° C. on a laboratory rocker for 30 minutes. Compound 19 (200 mg, 0.404mmol, 1 eq) was then added to 20 mL of the SIF suspension and rocked at37° C. overnight. The suspension was added to a separatory funnel anddiluted with additional water (20 mL). Product was extracted withdichloromethane (40 mL) three times, then the organic layer was driedover magnesium sulfate. After filtering out the salts, the solution wasconcentrated by rotary evaporation and re-dissolved in DMSO beforeinjection and purification by reverse phase C18 column chromatography(gradient: 10% acetonitrile in deionized water to 100% acetonitrile).Fractions containing product were lyophilized to yield the titlecompound as a white powder (95 mg, 0.223 mmol, 55% yield). LCMS [M−H]⁻:424.2. ¹H NMR (400 MHz, DMSO-d₆) δ 6.84 (d, J=8.7 Hz, 1H), 6.76 (d,J=2.8 Hz, 1H), 6.60 (dd, J=8.7, 2.9 Hz, 1H), 5.41 (d, J=5.3 Hz, 1H),5.02-4.93 (m, 2H), 4.84 (dd, J=9.7, 7.8 Hz, 1H), 3.83 (dd, J=11.3, 5.5Hz, 1H), 3.71-3.59 (m, 1H), 3.38 (t, J=10.8 Hz, 1H), 2.33-2.08 (m, 4H),1.56-1.40 (m, 4H), 0.89-0.77 (m, 6H).

Compound 33:5-amino-2-(((2S,3R,4S,5R)-3,4,5-triacetoxytetrahydro-2H-pyran-2-yl)oxy)benzoicacid

(3R,4S,5R)-tetrahydro-2H-pyran-2,3,4,5-tetrayl tetraacetate (3.90 g,12.3 mmol) was dissolved in tetrahydrofuran (80.0 mL). The solution wasstirred at room temperature when methylamine (40% wt. in water, 1.59 mL,18.4 mmol) was added dropwise. The mixture was stirred overnight andconcentrated to give crude(3R,4S,5R)-2-hydroxytetrahydro-2H-pyran-3,4,5-triyl triacetate as abrown oil.

Crude (3R,4S,5R)-2-hydroxytetrahydro-2H-pyran-3,4,5-triyl triacetate(500 mg, 1.81 mmol) was dissolved in N N-dimethylformamide (4.00 mL) atroom temperature. The solution was stirred when benzyl2-fluoro-5-nitrobenzoate (752 mg, 2.73 mmol) and then1,4-diazabicyclo[2.2.2]octane (1.06 g, 9.36 mmol) were added. Stirringwas continued for 90 h. Water (50 mL) was added and the mixture wasextracted with ethyl acetate (5×20 mL). The combined organic layers werewashed with water (20 mL), brine (2×20 mL), dried over anhydrous Na₂SO₄,filtered and concentrated to give a brown oil. The crude material wasadsorbed on celite and purified by automated chromatography (40 g, SiO₂,0 to 60% ethyl acetate in hexanes) to afford(2S,3R,4S,5R)-2-(2-((benzyloxy)carbonyl)-4-nitrophenoxy)tetrahydro-2H-pyran-3,4,5-triyltriacetate (413 mg, 43%) as a pale yellow foam. LCMS calcd for C₂₅H₂₅O₁₂531.14, found 554.4 [M+Na] at 1.86 min.

(2S,3R,4S,5R)-2-(2-((benzyloxy)carbonyl)-4-nitrophenoxy)tetrahydro-2H-pyran-3,4,5-triyltriacetate (413 mg, 0.777 mmol) was dissolved in methanol (3.00 mL) atroom temperature. The solution was stirred under nitrogen when palladiumon carbon (10% wt., 50.0 mg, 0.0470 mmol) was added. The mixture wasdegassed with hydrogen and then stirred under hydrogen overnight. Themixture was filtered on celite and concentrated to give a brown oil. Thecrude material was purified by automated reverse phase chromatography(24 g, C18, 5 to 40% acetonitrile in 10 mM aqueous ammonium formate) asa solution in N N-dimethylformamide (10% water). After lyophilization,5-amino-2-(((2S,3R,4S,5R)-3,4,5-triacetoxytetrahydro-2H-pyran-2-yl)oxy)benzoicacid (59.8 mg, 19%) was obtained as a white solid. ¹H NMR (400 MHz,DMSO-d₆) δ 6.87 (d, J=8.8 Hz, 1H), 6.79 (d, J=2.9 Hz, 1H), 6.62 (dd,J=8.7, 2.9 Hz, 1H), 5.21 (t, J=8.8 Hz, 1H), 5.11 (d, J=7.0 Hz, 1H), 4.96(dd, J=9.0, 7.0 Hz, 1H), 4.89 (td, J=8.9, 5.2 Hz, 1H), 4.04 (dd, J=11.6,5.2 Hz, 1H), 3.62 (dd, J=11.6, 9.1 Hz, 1H), 2.02-1.97 (m, 9H). LCMScalcd for C₁₈H₂₁O₁₀ 411.12, found 410.3 [M−H] at 1.10 min.

Compound 34:5-amino-2-(((2R,3R,4R,5R)-3,4,5-triacetoxytetrahydro-2H-pyran-2-yl)oxy)benzoicacid

D-(−)-ribose (1.99 g, 13.1 mmol) was dissolved in pyridine (40 mL) undernitrogen. The solution was stirred when acetic acid (10.0 mL, 106 mmol)and then 4-dimethylaminopyridine (126 mg, 1.01 mmol) were added.Stirring was continued overnight Water (150 mL) was added and after 1 hof additional stirring, the mixture was extracted with ethyl acetate(3×40 mL). The combined organic layers were washed with saturatedaqueous NaHCO₃ (3×50 mL), water (50 mL), brine (2×50 mL), dried overanhydrous Na₂SO₄, filtered and concentrated to give a pale yellow oil.The crude material was adsorbed on celite and purified by automatedchromatography (80 g, SiO₂, 0 to 80% ethyl acetate in heptanes) toafford (3R,4R,5R)-tetrahydro-2H-pyran-2,3,4,5-tetrayl tetraacetate (3.90g, 93%) as a colorless oil.(3R,4R,5R)-tetrahydro-2H-pyran-2,3,4,5-tetrayl tetraacetate (3.6 g, 11.3mmol) was dissolved in acetonitrile (14.0 mL) at room temperature.Aqueous perchloric acid (70% wt., 974 μL, 11.3 mmol) was added in oneportion and the mixture was stirred for 1 h. The mixture was washed withaqueous saturated NaHCO₃, water and brine, dried over anhydrous Na₂SO₄,filtered and concentrated to give crude(3R,4R,5R)-2-hydroxytetrahydro-2H-pyran-3,4,5-triyl triacetate (1.11 g,36%). ¹H NMR (400 MHz, DMSO-d₆) δ 7.02 (s, 1H, OH), 5.93-5.82 (m, 1H),5.51-5.04 (m, 1H), 4.99-4.44 (m, 2H), 3.98-3.38 (m, 2H), 2.12-1.91 (m,9H).

(3R,4R,5R)-2-hydroxytetrahydro-2H-pyran-3,4,5-triyl triacetate (1.11 g,4.02 mmol) was dissolved in N N-dimethylformamide (7.0 mL). The solutionwas stirred at room temperature when benzyl 2-fluoro-5-nitrobenzoate(1.11 g, 4.02 mmol) and then 1,4-diazabicyclo[2.2.2]octane (2.28 g, 20.1mmol) were added. Stirring was continued for 2 d. Then, water was added.The aqueous layer was extracted with ethyl acetate. The combined organiclayers were washed with brine, dried over anhydrous Na₂SO₄, filtered andconcentrated to give a brown oil. The crude material was adsorbed oncelite and purified by automated chromatography (SiO₂, ethyl acetategradient in hexanes) to give(3R,4R,5R)-2-(2-((benzyloxy)carbonyl)-4-nitrophenoxy)tetrahydro-2H-pyran-3,4,5-triyltriacetate (295 mg, 14%) as a yellow oil. ¹H NMR (400 MHz, CDCl₃) δ 8.70(d, J=2.9 Hz, 1H), 8.31 (dd, J=9.2, 2.9 Hz, 1H), 7.44-7.27 (m, 6H), 6.00(d, J=4.8 Hz, 0.5 Ha), 5.73 (d, J=2.7 Hz, 1 Ha′), 5.46 (t, J=3.6 Hz,1H), 5.38 (app q, J=12.1 Hz, 2H), 5.29-5.26 (m, 1H), 5.15-4.99 (m, 2H),4.02-3.94 (m, 2H), 3.80 (dd, J=12.9, 3.1 Hz, 1H), 2.20-1.97 (m, 9H).LCMS calcd for C₂₅H₂₅NO₁₂ 531.14, found 554.3 [M+Na] at 1.79 min.

(3R,4R,5R)-2-(2-((benzyloxy)carbonyl)-4-nitrophenoxy)tetrahydro-2H-pyran-3,4,5-triyltriacetate (280 mg, 527 μmol) was dissolved in methanol (5.0 mL).Palladium on carbon (10% wt., 22.4 mg, 21.1 μmol) was added to thestirring solution under nitrogen. The mixture was degassed with hydrogenand allowed to stir under hydrogen for 2 h. The mixture was filteredthrough a pad of celite and washed with methanol and dichloromethane.The filtrate was concentrated and purified by automated reverse phasechromatography (C18, 15 to 25% acetonitrile in 10 mM aqueous ammoniumformate). After lyophilisation,5-amino-2-(((2R,3R,4R,5R)-3,4,5-triacetoxytetrahydro-2H-pyran-2-yl)oxy)benzoicacid (26.7 mg, 12%) as a white solid. ¹H NMR (400 MHz, DMSO-d₆) δ 6.85(dd, J=5.6, 2.7 Hz, 2H), 6.60 (dd, J=8.6, 2.5 Hz, 1H), 5.36 (t, J=3.6Hz, 1H), 5.31 (d, J=3.0 Hz, 1H), 5.15 (t, J=3.3 Hz, 1H), 5.09 (d, J=3.2Hz, 1H), 4.18 (dd, J=12.8, 1.9 Hz, 1H), 3.73 (dd, J=12.7, 3.7 Hz, 1H),2.04 (d, J=4.3 Hz, 7H), 1.93 (s, 3H). LCMS calcd for C₁₈H₂₁NO₁₀ 411.12,found 412.1 [M+H] at 1.02 min

Compound35:5-amino-2-(((2R,3R,4R,5S,6S)-3,4,5-tris(butyryloxy)-6-methyltetrahydro-2H-pyran-2-yl)oxy)benzoicacid

(3R,4R,5S,6S)-2-hydroxy-6-methyltetrahydro-2H-pyran-3,4,5-triyltributyrate (316 mg, 0.844 mmol), benzyl 2-hydroxy-5-nitrobenzoate (355mg, 1.30 mmol) and triphenylphosphine (374 mg, 1.43 mmol) were dissolvedin dry tetrahydrofuran (2.50 mL) under nitrogen at room temperature. Thesolution was stirred at 0° C. when di-tert-butyl azodicarboxylate (299mg, 1.27 mmol) was added in one portion. After 30 min of additionalstirring, the flask was removed from the cooling bath, the mixture wasallowed to warm up till room temperature and stir overnight. The mixturewas adsorbed on celite and purified by automated chromatography (40 g,SiO₂, 0 to 30% ethyl acetate in hexanes) to afford(2R,3R,4R,5S,6S)-2-(2-((benzyloxy)carbonyl)-4-nitrophenoxy)-6-methyltetrahydro-2H-pyran-3,4,5-triyltributyrate (197 mg, 37%). LCMS calcd for C₃₂H₃₉NO₁₂ 629.25, found 647.0[M+NH₄] at 2.20 min.

(2R,3R,4R,5S,6S)-2-(2-((benzyloxy)carbonyl)-4-nitrophenoxy)-6-methyltetrahydro-2H-pyran-3,4,5-triyltributyrate (197 mg, 0.313 mmol) was dissolved in methanol (2.50 mL) atroom temperature. The solution was stirred under nitrogen when palladiumon carbon (10% wt., 2.30 mg, 0.0216 mmol) was added in one portion. Thesuspension was stirred, degassed with hydrogen and allowed to stir underhydrogen overnight. The mixture was diluted with dichloromethane andfiltered on celite. The crude material was concentrated and purified byautomated reverse phase chromatography (12 g, C18, 10 to 60%acetonitrile in 10 mM aqueous ammonium formate) as a solution in NN-dimethylformamide (10% water). After lyophilisation,5-amino-2-(((2R,3R,4R,5S,6S)-3,4,5-tris(butyryloxy)-6-methyltetrahydro-2H-pyran-2-yl)oxy)benzoicacid (78.4 mg, 49%) was obtained as a white solid. ¹H NMR (400 MHz,DMSO-d₆) δ 6.89 (d, J=2.9 Hz, 1H), 6.87 (d, J=8.7 Hz, 1H), 6.65 (dd,J=8.7, 2.9 Hz, 1H), 5.58 (dd, J=3.3, 0.9 Hz, 1H), 5.30 (d, J=1.0 Hz,1H), 5.18 (dd, J=10.2, 3.4 Hz, 1H), 5.05 (brs, 2H), 4.90 (t, J=9.9 Hz,1H), 3.80-3.70 (m, J=6.2 Hz, 1H), 2.41-2.34 (m, 2H), 2.33-2.22 (m, 2H),2.14 (td, J=7.2, 3.2 Hz, 2H), 1.66-1.56 (m, 2H), 1.56-1.42 (m, 4H), 1.12(d, J=6.2 Hz, 3H), 0.94 (t, J=7.4 Hz, 3H), 0.89-0.80 (m, 6H). LCMS calcdfor C₂₅H₃₅NO₁₀ 509.23, found 508.3 [M−H] at 1.74 min.

Compound 36:5-amino-2-(((2S,3R,4S,5R,6R)-3,4,5-tris(butyryloxy)-6-((butyryloxy)methyl)tetrahydro-2H-pyran-2-yl)oxy)benzoicacid

2R,3R,4S,5R)-2-((butyryloxy)methyl)-6-hydroxytetrahydro-2H-pyran-3,4,5-triyltributyrate (239 mg, 519 μmol) was dissolved in N N-dimethylformamide (1mL). The solution was stirred at room temperature when benzyl2-fluoro-5-nitrobenzoate (186 mg, 675 μmol) and then1,4-diazabicyclo[2.2.2]octane (294 mg, 2.59 mmol) were added. Stirringwas continued for 2 d and water was added. The aqueous layer wasextracted with ethyl acetate. The combined organic layers were washedwith brine, dried over anhydrous Na₂SO₄, filtered and concentrated togive a brown oil. The crude material was adsorbed on celite to bepurified by automated chromatography (SiO₂, ethyl acetate gradient inhexanes) to afford(3R,4S,5R,6R)-2-(2-((benzyloxy)carbonyl)-4-nitrophenoxy)-6-((butyryloxy)methyl)tetrahydro-2H-pyran-3,4,5-triyltributyrate (169 mg, 45%). LCMS calcd for C₃₆H₄₅NO₁₄ 715.28, found 733.6[M+NH₄] at 2.26 min.

(3R,4S,5R,6R)-2-(2-((benzyloxy)carbonyl)-4-nitrophenoxy)-6-((butyryloxy)methyl)tetrahydro-2H-pyran-3,4,5-triyltributyrate (169 mg, 236 μmol) was dissolved in methanol (5.0 mL) atroom temperature. The solution was stirred under nitrogen when palladiumon carbon (10% wt., 25.1 mg, 23.6 μmol) was added in one portion. Then,the solvent was degassed with hydrogen and the reaction was allowed tostir under hydrogen for 2 h. The mixture was diluted withdichloromethane and filtered on celite. The crude material was purifiedby automated reverse phase chromatography (C18, 25% to 65% acetonitrilein 10 mM aqueous ammonium formate). After lyophilisation,5-amino-2-(((2S,3R,4S,5R,6R)-3,4,5-tris(butyryloxy)-6-((butyryloxy)methyl)tetrahydro-2H-pyran-2-yl)oxy)benzoicacid (32.3 mg, 23%) was obtained as a white solid ¹H NMR (400 MHz,DMSO-d₆) δ 6.88 (d, J=8.8 Hz, 1H), 6.76 (d, J=2.8 Hz, 1H), 6.60 (dd,J=8.8, 2.8 Hz, 1H), 5.36 (t, J=9.6 Hz, 1H), 5.21 (d, J=8.0 Hz, 1H),5.03-4.93 (m, 3H), 4.18-4.05 (m, 2H, H), 2.30-2.09 (m, 8H), 1.58-1.39(m, 8H), 0.90-0.77 (m, 12H). LCMS calcd for C₂₉H₄₁NO₁₂ 595.26, found613.3 [M+NH₄].

Compound 37:5-amino-2-(((2S,3R,4S,5R)-3,4,5-tris((3-(1H-indol-3-yl)propanoyl)oxy)tetrahydro-2H-pyran-2-yl)oxy)benzoicacid

3-Indolepropionic acid (20.0 g, 104 mmol) and dicyclohexylcarbodiimide(10.3 g, 49.3 mmol) were dissolved in tetrahydrofuran (345 mL). Thereaction was stirred under nitrogen for 2 d. The solution was filtered,washed with tetrahydrofuran and the filtrate was concentrated to givecrude 3-(1H-indol-3-yl)propanoic anhydride (26 g, 69%).

Crude 3-(1H-indol-3-yl)propanoic anhydride (26.0 g, 72.1 mmol) wasdissolved in pyridine (150 mL) under nitrogen. 4-dimethylaminopyridine(450 mg, 3.61 mmol) and d-(+)-xylose (1.11 g, 7.43 mmol) were added. Themixture was stirred for 24 h. 1 N aqueous hydrochloric acid was addedand the aqueous layer was extracted with ethyl acetate. The combinedorganic layers were concentrated. The crude material was purified byautomated reverse phase chromatography (C18, 60 to 65% acetonitrile in10 mM aqueous ammonium formate) to afford(3R,4S,5R)-tetrahydro-2H-pyran-2,3,4,5-tetrayltetrakis(3-(1H-indol-3-yl)propanoate (3.49 g, 58%) as a yellowsuspension. LCMS calcd for C₄₉H₄₆N₄O₉ 834.33, found 833.6 [M−H] at 2.05min.

(3R,4S,5R)-tetrahydro-2H-pyran-2,3,4,5-tetrayltetrakis(3-(1H-indol-3-yl)propanoate) (1.06 g, 1.27 mmol) was dissolvedin acetonitrile (13.0 mL) at room temperature. Aqueous perchloric acid(70% wt., 110 μL, 1.27 mmol) was added and the mixture was stirred for 3h. The mixture was washed with saturated aqueous NaHCO₃, water and brineand dried over anhydrous Na₂SO₄, filtered and concentrated. The crudematerial was purified by automated reverse phase chromatography (C18,acetonitrile in 10 mM aqueous ammonium formate). After lyophilisation,(3R,4S,5R)-2-hydroxytetrahydro-2H-pyran-3,4,5-triyltris(3-(1H-indol-3-yl)propanoate (121 mg, 14%) was obtained as a yellowsolid. ¹H NMR (400 MHz, DMSO-d₆) δ 10.75 (s, 3H), 7.46-7.34 (m, 3H),7.25 (d, J=10.6 Hz, 3H), 7.02 (m, 8H), 6.94-6.82 (m, 4H), 5.41 (t, J=9.8Hz, 0.5H), 5.26 (s, 0.3H), 5.16 (s, 1H), 4.92-4.69 (m, 2H), 3.64 (m,2H), 2.84 (dd, J=15.8, 7.8 Hz, 6H), 2.53-2.49 (m, 6H). LCMS calcd forC₃₈H₃₇N₃O₈ 663.26, found 681.2 [M+NH₄] at 1.80 min.

(3R,4S,5R)-2-hydroxytetrahydro-2H-pyran-3,4,5-triyltris(3-(1H-indol-3-yl)propanoate) (121 mg, 182 μmol) was dissolved in NN-dimethylformamide (600 μL). The solution was stirred at roomtemperature when benzyl 2-fluoro-5-nitrobenzoate (65.2 mg, 237 μmol) andthen 1,4-diazabicyclo[2.2.2]octane (102 mg, 912 μmol) were added.Stirring was continued for 2 d. Then, water was added. The aqueous layerwas extracted with ethyl acetate. The combined organic layers werewashed with brine, dried over anhydrous Na₂SO₄, filtered andconcentrated to give a brown oil. The crude material was adsorbed oncelite and purified by automated chromatography (SiO₂, ethyl acetategradient in hexanes) to afford(3R,4S,5R)-2-(2-((benzyloxy)carbonyl)-4-nitrophenoxy)tetrahydro-2H-pyran-3,4,5-triyltris(3-(1H-indol-3-yl)propanoate) (36.0 mg, 21%) as a yellow oil. LCMScalcd for C₅₂H₄₆N₄O₁₂ 918.31, found 936 [M+NH₄] at 2.10 min.

(3R,4S,5R)-2-(2-((benzyloxy)carbonyl)-4-nitrophenoxy)tetrahydro-2H-pyran-3,4,5-triyltris(3-(1H-indol-3-yl)propanoate) (36.0 mg, 39.2 μmol) was dissolved inmethanol (800 μL) and stirred at room temperature under nitrogen. Tothis stirring solution, palladium on carbon (10% wt. 4.17 mg, 3.92 μmol)was added. The suspension was degassed with hydrogen and allowed to stirunder hydrogen for 2 h. The mixture was filtered through celite andwashed with methanol. The filtrate was concentrated under vacuo. Thecrude material was purified by preparative HPLC-MS (CSH column,acetonitrile in 10 mM aqueous ammonium formate). After lyophilisation,3-carboxy-4-(((2S,3R,4S,5R)-3,4,5-tris((3-(1H-indol-3-yl)propanoyl)oxy)tetrahydro-2H-pyran-2-yl)oxy)benzenaminiumformate (4.20 mg, 13%) was obtained as a white solid. ¹H NMR (400 MHz,DMSO-d₆) δ 10.77 (t, J=12.5 Hz, 3H), 8.40 (s, 3H), 7.53-7.38 (m, 3H),7.28 (dd, J=8.0, 4.8 Hz, 3H), 7.03 (dd, J=13.2, 4.9 Hz, 6H), 6.96-6.85(m, 3H), 6.82-6.67 (m, 2H), 6.53 (d, J=7.0 Hz, 1H), 5.29 (t, J=8.5 Hz,1H), 5.18 (d, J=6.7 Hz, 1H), 5.06-5.01 (m, 1H), 4.91 (dd, J=13.5, 8.3Hz, 1H), 3.98 (dd, J=11.6, 4.9 Hz, 1H), 3.60-3.51 (m, 1H), 2.91-2.79 (m,7H), 2.74-2.67 (m, 1H), 2.63-2.54 (m, 4H). LCMS calcd for C₄₆H₄₂N₄O₁₀798.29, found 816 [M+NH₄] at 1.80 min.

Compound 38:2-(((2R,3R,4R,5S,6S)-3,4,5-tris(butyryloxy)-6-methyltetrahydro-2H-pyran-2-yl)oxy)benzoicacid

(3R,4R,5S,6S)-2-hydroxy-6-methyltetrahydro-2H-pyran-3,4,5-triyltributyrate (1.20 g, 3.20 mmol), benzyl 2-hydroxybenzoate (1.10 g, 4.81mmol) and triphenylphosphine (1.27 g, 4.81 mmol) were dissolved intetrahydrofuran (54.0 mL) and stirred at 0° C. Di-tert-butylazodicarboxylate (1.11 g, 4.81 mmol) was added portion wise and thereaction mixture was stirred at 0° C. for 1 h and allowed to warm uptill room temperature to stir overnight. The mixture was adsorbed onsilica to be purified by automated chromatography (100 g, SiO₂, 0 to 35%ethyl acetate in hexanes).(2S,3R,4R,5S,6S)-2-(2-((benzyloxy)carbonyl)phenoxy)-6-methyltetrahydro-2H-pyran-3,4,5-triyltributyrate (165 mg, 8.1%) and

(2R,3R,4R,5S,6S)-2-(2-((benzyloxy)carbonyl)phenoxy)-6-methyltetrahydro-2H-pyran-3,4,5-triyltributyrate (427 mg, 21%) were separated but containing otherimpurities.(2S,3R,4R,5S,6S)-2-(2-((benzyloxy)carbonyl)phenoxy)-6-methyltetrahydro-2H-pyran-3,4,5-triyltributyrate (166 mg, 284 μmol) was dissolved in methanol (9.00 mL) andstirred at room temperature under nitrogen. To this mixture was addedpalladium on carbon (10% wt., 11.0 mg, 73.0 μmol). The suspension wasdegassed with hydrogen and stir under hydrogen overnight. The mixturewas filtered through celite and washed with methanol. The filtrate wasconcentrated under vacuo. The crude material was purified by automatedchromatography (25 g, SiO₂, 0 to 100% ethyl acetate in hexanes) as asolution in dichloromethane to afford2-(((2S,3R,4R,5S,6S)-3,4,5-tris(butyryloxy)-6-methyltetrahydro-2H-pyran-2-yl)oxy)benzoicacid (9 mg, 6%) as a white solid. ¹H NMR (400 MHz, DMSO-d₆) δ 7.71 (d,J=8.2 Hz, 1H), 7.51 (t, J=8.1 Hz, 1H), 7.29 (d, J=8.0 Hz, 1H), 7.14 (t,J=7.9 Hz, 1H), 5.76 (d, J=3.1 Hz, 2H), 5.43 (d, J=8.3 Hz, 2H), 5.02 (t,J=9.8 Hz, 1H), 1.54 (ddt, J=22.7, 14.8, 7.4 Hz, 6H), 1.05 (d, J=6.3 Hz,3H), 0.95 (t, J=7.4 Hz, 3H), 0.85 (q, J=7.4 Hz, 6H). LCMS calcd forC₂₅H₃₄O₁₀ 494.22, found 512.3 [M+NH₄] at 1.94 min.

(2R,3R,4R,5S,6S)-2-(2-((benzyloxy)carbonyl)phenoxy)-6-methyltetrahydro-2H-pyran-3,4,5-triyltributyrate (427 mg, 711 μmol) was dissolved methanol (9.00 mL) andstirred at room temperature under nitrogen. To this mixture was addedpalladium on carbon (10% wt., 11.0 mg, 73.0 μmol). The suspension wasdegassed with hydrogen and stir under hydrogen overnight. The mixturewas filtered through celite and washed with methanol. The filtrate wasconcentrated under vacuo. The crude material was purified by automatedchromatography (50 g, SiO₂, 0 to 100% ethyl acetate in hexanes) as asolution in dichloromethane to afford2-(((2R,3R,4R,5S,6S)-3,4,5-tris(butyryloxy)-6-methyltetrahydro-2H-pyran-2-yl)oxy)benzoicacid (18 mg, 5%) as a white solid. ¹H NMR (400 MHz, DMSO-d₆) δ 7.62 (dd,J=7.5, 1.6 Hz, 1H), 7.46 (t, J=7.9 Hz, 1H), 7.14 (d, J=7.8 Hz, 1H), 7.08(td, 7.6, 0.8 Hz, 1H), 5.68 (d, J=0.8 Hz, 1H), 5.55 (dd, J=3.5, 1.0 Hz,1H), 5.22 (dd, J=10.2, 3.4 Hz, 1H), 4.90 (t, J=9.9 Hz, 1H), 3.90-3.82(m, 1H), 2.38 (t, J=7.1 Hz, 2H), 2.27 (m, 2H), 2.13 (td, J=7.2, 2.7 Hz,2H), 1.60 (m, 2H), 1.48 (m, 4H), 1.12 (d, J=6.2 Hz, 3H), 0.93 (t, J=7.4Hz, 3H), 0.85 (t, J=8.5 Hz, 3H), 0.83 (t, J=8.1 Hz, 3H). LCMS calcd forC₂₅H₃₄O₁₀ 494.22, found 493.3 [M−H] at 1.87 min.

Compound 39:5-amino-2-(((2S,3R,4R,5S,6S)-3,4,5-tris(butyryloxy)-6-methyltetrahydro-2H-pyran-2-yl)oxy)benzoicacid

(3R,4R,5S,6S)-2-hydroxy-6-methyltetrahydro-2H-pyran-3,4,5-triyltributyrate (197 mg, 0.526 mmol) and benzyl 2-fluoro-5-nitrobenzoate(194 mg, 0.705 mmol) were dissolved in N N-dimethylformamide (1.0 mL) atroom temperature. The solution was stirred when1,4-diazabicyclo[2.2.2]octane (282 mg, 2.51 mmol) was added in oneportion and stirring was continued for 88 h. Water (60 mL) was added andthe aqueous layer was extracted with ethyl acetate (3×20 mL). Thecombined organic layers were washed with brine (3×20 mL), dried overanhydrous Na₂SO₄, filtered and concentrated. The crude material wasadsorbed on celite and purified by automated chromatography (12 g, SiO₂,0 to 20% ethyl acetate in hexanes) to afford(2S,3R,4R,5S,6S)-2-(2-((benzyloxy)carbonyl)-4-nitrophenoxy)-6-methyltetrahydro-2H-pyran-3,4,5-triyltributyrate (136 mg, 41%) as a colorless gum. ¹H NMR (400 MHz, CDCl₃) δ8.75 (d, J=2.9 Hz, 1H), 8.33 (dd, J=9.2, 2.9 Hz, 1H), 7.51-7.46 (m, 2H),7.41-7.30 (m, 4H), 5.66 (d, J=1.9 Hz, 1H), 5.61 (dd,0=10.2, 3.5 Hz, 1H),5.58-5.44 (m, 3H), 5.23 (t, J=10.0 Hz, 1H), 3.99-3.91 (m, 1H), 2.51-2.38(m, 2H), 2.31-2.20 (m, 4H), 1.77-1.67 (m, 2H), 1.67-1.57 (m, 4H), 1.18(d, J=6.3 Hz, 3H), 1.01 (t, J=7.4 Hz, 3H), 0.97-0.90 (m, 6H). LCMS calcdfor C₃₂H₃₉NO₁₂ 629.25, found 647.1 [M+NH₄] at 2.25 min.

(2S,3R,4R,5S,6S)-2-(2-((benzyloxy)carbonyl)-4-nitrophenoxy)-6-methyltetrahydro-2H-pyran-3,4,5-triyltributyrate (136 mg, 0.216 mmol) was dissolved in methanol (2.00 mL) atroom temperature under nitrogen. Palladium on carbon (10% wt., 2.30 mg,0.0216 mmol) was added in one portion. The suspension was stirred,degassed with hydrogen and allowed to stir under hydrogen overnight. Themixture was diluted with dichloromethane and filtered on celite. Thefiltrate was concentrated and this material was purified by preparativeHPLC-MS (CSH column, 40 to 60% acetonitrile in 10 mM aqueous ammoniumformate) as a solution in N N-dimethylformamide (10% water). Afterlyophilization,5-amino-2-(((2S,3R,4R,5S,6S)-3,4,5-tris(butyryloxy)-6-methyltetrahydro-2H-pyran-2-yl)oxy)benzoicacid (70.9 mg, 64%) was obtained as a white solid. ¹H NMR (400 MHz,DMSO-d₆) δ 6.93 (d, J=2.9 Hz, 1H), 6.90 (d, J=8.7 Hz, 1H), 6.66 (dd,J=8.7, 2.9 Hz, 1H), 5.45 (dd, J=3.4, 1.8 Hz, 1H), 5.32 (dd, J=10.2, 3.4Hz, 1H), 5.30 (d, J=1.4 Hz, 1H), 5.21-5.03 (m, 2H), 5.00 (t, J=10.1 Hz,1H), 4.20-4.12 (m, 1H), 2.40-2.22 (m, 4H), 2.15 (td, J=7.2, 1.3 Hz, 2H),1.64-1.43 (m, 6H), 1.08 (d, J=6.3 Hz, 3H), 0.93 (t, J=7.4 Hz, 3H),0.89-0.81 (m, 6H). LCMS calcd for C₂₅H₃₅NO₁₀ 509.23, found 508.3 [M−H]at 1.78 min.

Compound 40:2-(((2S,3R,4R,5S,6S)-3,4,5-tris(butyryloxy)-6-methyltetrahydro-2H-pyran-2-yl)oxy)benzoicacid

(3R,4R,5S,6S)-2-hydroxy-6-methyltetrahydro-2H-pyran-3,4,5-triyltributyrate (1.20 g, 3.20 mmol), benzyl 2-hydroxybenzoate (1.10 g, 4.81mmol) and triphenylphosphine (1.27 g, 4.81 mmol) were dissolved intetrahydrofuran (54.0 mL) and stirred at 0° C. Di-tert-butylazodicarboxylate (1.11 g, 4.81 mmol) was added portion wise and thereaction mixture was stirred at 0° C. for 1 h and allowed to warm uptill room temperature to stir overnight. The mixture was adsorbed onsilica to be purified by automated chromatography (100 g, SiO₂, 0 to 35%ethyl acetate in hexanes).(2S,3R,4R,5S,6S)-2-(2-((benzyloxy)carbonyl)phenoxy)-6-methyltetrahydro-2H-pyran-3,4,5-triyltributyrate (165 mg, 8.1%) and(2R,3R,4R,5S,6S)-2-(2-((benzyloxy)carbonyl)phenoxy)-6-methyltetrahydro-2H-pyran-3,4,5-triyltributyrate (427 mg, 21%) were separated but containing otherimpurities.

(2S,3R,4R,5S,6S)-2-(2-((benzyloxy)carbonyl)phenoxy)-6-methyltetrahydro-2H-pyran-3,4,5-triyltributyrate (166 mg, 284 μmol) was dissolved in methanol (9.00 mL) andstirred at room temperature under nitrogen. To this mixture was addedpalladium on carbon (10% wt., 11.0 mg, 73.0 μmol). The suspension wasdegassed with hydrogen and stir under hydrogen overnight. The mixturewas filtered through celite and washed with methanol. The filtrate wasconcentrated under vacuo. The crude material was purified by automatedchromatography (25 g, SiO₂, 0 to 100% ethyl acetate in hexanes) as asolution in dichloromethane to afford2-(((2S,3R,4R,5S,6S)-3,4,5-tris(butyryloxy)-6-methyltetrahydro-2H-pyran-2-yl)oxy)benzoicacid (9 mg, 6%) as a white solid. 1H NMR (400 MHz, DMSO-d6) δ 7.71 (d,J=8.2 Hz, 1H), 7.51 (t, J=8.1 Hz, 1H), 7.29 (d, J=8.0 Hz, 1H), 7.14 (t,J=7.9 Hz, 1H), 5.76 (d, J=3.1 Hz, 2H), 5.43 (d, J=8.3 Hz, 2H), 5.02 (t,J=9.8 Hz, 1H), 1.54 (ddt, J=22.7, 14.8, 7.4 Hz, 6H), 1.05 (d, J=6.3 Hz,3H), 0.95 (t, J=7.4 Hz, 3H), 0.85 (q, J=7.4 Hz, 6H). LCMS calcd forC25H34O10 494.22, found 512.3 [M+NH4] at 1.94 min.

(2R,3R,4R,5S,6S)-2-(2-((benzyloxy)carbonyl)phenoxy)-6-methyltetrahydro-2H-pyran-3,4,5-triyltributyrate (427 mg, 711 μmol) was dissolved methanol (9.00 mL) andstirred at room temperature under nitrogen. To this mixture was addedpalladium on carbon (10% wt., 11.0 mg, 73.0 μmol). The suspension wasdegassed with hydrogen and stir under hydrogen overnight. The mixturewas filtered through celite and washed with methanol. The filtrate wasconcentrated under vacuo. The crude material was purified by automatedchromatography (50 g, SiO₂, 0 to 100% ethyl acetate in hexanes) as asolution in dichloromethane to afford2-(((2R,3R,4R,5S,6S)-3,4,5-tris(butyryloxy)-6-methyltetrahydro-2H-pyran-2-yl)oxy)benzoicacid (18 mg, 5%) as a white solid. 1H NMR (400 MHz, DMSO-d6) δ 7.62 (dd,J=7.5, 1.6 Hz, 1H), 7.46 (t, J=7.9 Hz, 1H), 7.14 (d, J=7.8 Hz, 1H), 7.08(td, J=7.6, 0.8 Hz, 1H), 5.68 (d, J=0.8 Hz, 1H), 5.55 (dd, J=3.5, 1.0Hz, 1H), 5.22 (dd, J=10.2, 3.4 Hz, 1H), 4.90 (t, J=9.9 Hz, 1H),3.90-3.82 (m, 1H), 2.38 (t, J=7.1 Hz, 2H), 2.27 (m, 2H), 2.13 (td,J=7.2, 2.7 Hz, 2H), 1.60 (m, 2H), 1.48 (m, 4H), 1.12 (d, J=6.2 Hz, 3H),0.93 (t, J=7.4 Hz, 3H), 0.85 (t, J=8.5 Hz, 3H), 0.83 (t, J=8.1 Hz, 3H).LCMS calcd for C25H34O10 494.22, found 493.3 [M−H].

Compound 41:5-amino-2-(((3S,4R,5R,6S)-3,4,5-tris(butyryloxy)-6-methyltetrahydro-2H-pyran-2-yl)oxy)benzoicacid

(3S,4R,5R,6S)-2-hydroxy-6-methyltetrahydro-2H-pyran-3,4,5-triyltributyrate (500 mg, 1.34 mmol), tert-butyl5-((tert-butoxycarbonyl)amino)-2-hydroxybenzoate (620 mg, 2.00 mmol) andtriphenylphosphine (531 mg, 2.00 mmol) were dissolved in tetrahydrofuran(22.0 mL) and stirred at 0° C. Di-tert-butyl azodicarboxylate (471 mg,2.00 mmol) was added and stirring was continued at 0° C. for 1 h, thenat room temperature overnight. The reaction mixture was concentrated.The crude material was purified by automated chromatography (SiO₂, 30%ethyl acetate in hexanes) to afford(3S,4R,5R,6S)-2-(2-(tert-butoxycarbonyl)-4-((tert-butoxycarbonyl)amino)phenoxy)-6-methyltetrahydro-2H-pyran-3,4,5-triyltributyrate (213 mg, 24%). [M+NH₄ ⁺]=683, room temperature=2.21. ¹H NMR(400 MHz, CDCl₃) δ 7.58-6.95 (m, 3H), 5.48 (dd, J=10.5, 8.0 Hz, 1H),5.28 (d, J=3.4 Hz, 1H), 5.10 (dd, J=10.4, 3.4 Hz, 1H), 5.00 (d, J=8.0Hz, 1H), 3.88 (q, J=6.5 Hz, 1H), 2.55-2.15 (m, 2H), 1.76-1.56 (m, 3H),1.55 (s, 4H), 1.49 (s, 3H), 1.46 (s, 2H), 1.26-1.14 (m, 2H), 0.99 (t,J=7.4 Hz, 1H), 0.89 (t, J=7.4 Hz, 2H).

(3S,4R,5R,6S)-2-(2-(tert-butoxycarbonyl)-4-((tert-butoxycarbonyl)amino)phenoxy)-6-methyltetrahydro-2H-pyran-3,4,5-triyltributyrate (200 mg, 300 μmol) was dissolved in dichloromethane (2.00mL) at 0° C. To the solution was added hydrochloric acid (4 M in1,4-dioxane, 158 μL, 631 μmol). The resulting mixture was stirred at 0°C. for 30 min, then at room temperature for 2 h. Solvents wereevaporated and the residue was purified by automated reverse phasechromatography (C18, acetonitrile in 10 mM aqueous ammonium formate)After lyophilisation,5-amino-2-(((3S,4R,5R,6S)-3,4,5-tris(butyryloxy)-6-methyltetrahydro-2H-pyran-2-yl)oxy)benzoicacid (10.0 mg, 13%) was obtained as a white solid. ¹H NMR (400 MHz,DMSO-d₆) δ 6.86 (d, J=9.1 Hz, 1H), 6.74 (d, J=2.9 Hz, 1H), 6.60 (dd,J=9.8, 2.1 Hz, 1H), 5.21-5.06 (m, 4H), 5.01-4.91 (m, 1H), 2.67-2.63 (m,1H), 2.53 (s, 1H), 2.45-2.07 (m, 7H), 1.60 (dt, J=14.6, 7.3 Hz, 2H),1.52-1.39 (m, 4H), 1.07 (d, J=6.4 Hz, 3H), 0.92 (t, J=7.4 Hz, 3H), 0.80(t, J=7.4 Hz, 6H). LCMS calcd for C₂₅H₃₅NO₁₀ 509.23, found 508.2 [M−H]at 1.68 min.

Compound 42:2-(((3R,4R,5R)-3,4,5-tris(butyryloxy)tetrahydro-2H-pyran-2-yl)oxy)benzoicacid(3R,4R,5R)-2-hydroxytetrahydro-2H-pyran-3,4,5-triyl tributyrate (200mg, 555 μmol), benzyl 2-hydroxybenzoate (190 mg, 832 μmol) andtriphenylphosphine (221 mg, 832 μmol) were dissolved in tetrahydrofuran(4.0 mL) and stirred at 0° C. Di-tert-butyl azodicarboxylate (196 mg,832 μmol) was added and stirring continued at 0° C. for 1 h, then atroom temperature overnight. The reaction mixture was concentrated andpurified by automated chromatography (SiO₂, ethyl acetate gradient inhexanes) to afford(2R,3R,4R,5R)-2-(2-((benzyloxy)carbonyl)phenoxy)tetrahydro-2H-pyran-3,4,5-triyltributyrate (229 mg, 72%) as a colorless oil. ¹H NMR (400 MHz, CDCl₃) δ7.74 (dd, J=7.7, 1.8 Hz, 1H), 7.46-7.31 (m, 6H), 7.19 (d, J=8.3 Hz, 1H),7.08 (t, J=7.5 Hz, 1H), 5.57 (broad s, 1H), 5.51 (d, J=3.4 Hz, 1H), 5.31(dd, J=29.5, 12.5 Hz, 2H), 5.22 (broad s, 1H), 5.15-5.09 (m, 1H), 4.21(dd, J=11.3, 8.8 Hz, 1H), 3.65 (dd, J=11.3, 4.2 Hz, 1H), 2.50-2.26 (m,6H), 1.74-1.57 (m, 6H), 1.01-0.84 (m, 9H). LCMS calcd for C₃₁H₃₈O₁₀570.25, found 588.4 [M+NH₄] at 2.19 min.

(2R,3R,4R,5R)-2-(2-((benzyloxy)carbonyl)phenoxy)tetrahydro-2H-pyran-3,4,5-triyltributyrate (229 mg, 401 μmol) was dissolved in methanol (4.0 mL) atroom temperature. The solution was stirred under nitrogen when palladiumon carbon (10% wt., 42.7 mg, 40.1 μmol) was added in one portion. Themixture was degassed with hydrogen and allowed to stir under hydrogenfor 3 h. The mixture was diluted with dichloromethane and filtered oncelite. The crude material was adsorbed on celite and purified byautomated reverse phase chromatography (C18, acetonitrile in 10 mMaqueous ammonium formate). After lyophilisation,2-(((2R,3R,4R,5R)-3,4,5-tris(butyryloxy)tetrahydro-2H-pyran-2-yl)oxy)benzoicacid obtained as an oil (92.0 mg, 48%). ¹H NMR (400 MHz, DMSO-d₆) δ 7.61(dd, J=7.7, 1.7 Hz, 1H), 7.51-7.44 (m, 1H), 7.18 (d, J=8.4 Hz, 1H), 7.09(t, J=7.5 Hz, 1H), 5.63 (broad d, J=2.7 Hz, 1H), 5.45 (broad t, J=3.4Hz, 1H), 5.31 (broad t, J=2.9 Hz, 1H), 5.09 (broad dt, J=6.6, 3.5 Hz,1H), 4.00 (dd, J=12.0, 6.3 Hz, 1H), 3.81 (dd, J=12.0, 3.2 Hz, 1H),2.38-2.19 (m, 6H), 1.62-1.44 (m, 6H), 0.86 (td, J=7.4, 2.9 Hz, 9H).UP-LCMS calcd for C₂₄H₃₂O₁₀ 480.20, found 503.2 [M+Na] at 1.72 min.

Compound 43:5-amino-2-(((2R,3R,4S,5R,6R)-3,4,5-tris(butyryloxy)-6-((butyryloxy)methyl)tetrahydro-2H-pyran-2-yl)oxy)benzoicacid

Compound 43 was isolated as a minor isomer during the preparation ofcompound 36.

(2R,3R,4S,5R)-2-((butyryloxy)methyl)-6-hydroxytetrahydro-2H-pyran-3,4,5-triyltributyrate (239 mg, 519 μmol) was dissolved in N N-dimethylformamide (1mL). The solution was stirred at room temperature when benzyl2-fluoro-5-nitrobenzoate (186 mg, 675 μmol) and then1,4-diazabicyclo[2.2.2]octane (294 mg, 2.59 mmol) were added. Stirringwas continued for 2 d and water was added. The aqueous layer wasextracted with ethyl acetate. The combined organic layers were washedwith brine, dried over anhydrous Na₂SO₄, filtered and concentrated togive a brown oil. The crude material was adsorbed on celite to bepurified by automated chromatography (SiO₂, ethyl acetate gradient inhexanes) to afford(3R,4S,5R,6R)-2-(2-((benzyloxy)carbonyl)-4-nitrophenoxy)-6-((butyryloxy)methyl)tetrahydro-2H-pyran-3,4,5-triyltributyrate (169 mg, 45%). LCMS calcd for C₃₆H₄₅NO₁₄ 715.28, found 733.6[M+NH4] at 2.26 min.

(3R,4S,5R,6R)-2-(2-((benzyloxy)carbonyl)-4-nitrophenoxy)-6-((butyryloxy)methyl)tetrahydro-2H-pyran-3,4,5-triyltributyrate (169 mg, 236 μmol) was dissolved in methanol (5.0 mL) atroom temperature. The solution was stirred under nitrogen when palladiumon carbon (10% wt., 25.1 mg, 23.6 μmol) was added in one portion. Then,the solvent was degassed with hydrogen and the reaction was allowed tostir under hydrogen for 2 h. The mixture was diluted withdichloromethane and filtered on celite. The crude material was purifiedby automated reverse phase chromatography (C18, 25% to 65% acetonitrilein 10 mM aqueous ammonium formate). After lyophilisation,5-amino-2-(((2R,3R,4S,5R,6R)-3,4,5-tris(butyryloxy)-6-((butyryloxy)methyl)tetrahydro-2H-pyran-2-yl)oxy)benzoicacid (9 mg, 6%) was obtained as a white solid. ¹H NMR (400 MHz, DMSO-d₆)δ 6.82 (d, J=8.5 Hz, 1H), 6.79 (s, 1H), 6.58 (d, J=8.3 Hz, 1H), 5.56 (d,J=3.6 Hz, 1H), 5.52 (t, J=9.9 Hz, 1H), 5.04 (t, J=9.9 Hz, 1H), 4.93 (dd,J=10.3, 3.6 Hz, 1H), 4.38 (broad s, 1H), 4.10 (dd, J=12.4, 5.1 Hz, 1H),3.98 (dd, J=12.4, 2.0 Hz, 1H), 2.37-2.15 (m, 8H), 1.56-1.39 (m, 8H),0.90-0.73 (m, 12H). LCMS calcd for C₂₉H₄₁NO₁₂ 595.26, found 613.3[M+NH₄] at 1.84 min.

Compound 44:5-amino-2-(((2S,3R,4S,5R)-3,4,5-tris(butyryloxy)tetrahydro-2H-pyran-2-yl)oxy)benzoicacid hydrochloride

Xylose tributyrate (1.87 g, 5.18 mmol, 1 eq), tert-butyl5-((tert-butoxycarbonyl)amino)-2-hydroxybenzoate (2.4 g, 7.8 mmol, 1.5eq) and triphenylphosphine (2.05 g, 7.8 mmol, 1.5 eq) were dissolved inanhydrous THF (37.5 mL) under N₂ and cooled to 0° C. Di-tert-butylazodicarboxylate (1.8 g, 7.8 mmol, 1.5 eq) was added and the reactionwas stirred at 0° C. for 1 hour under N2. The ice bath was removed andthe reaction was stirred at room temperature overnight. The reactionmixture was loaded directly onto silica and dried by rotary evaporation.The solid loaded sample was purified by multiple rounds of columnchromatography (gradient: 0-50% ethyl acetate in hexanes) to separatethe anomers and yield(2S,3R,4S,5R)-2-(2-(tert-butoxycarbonyl)-4-((tert-butoxycarbonyl)amino)phenoxy)tetrahydro-2H-pyran-3,4,5-triyltributyrate (45 mg). The purified compound was dissolved in chloroform(1 mL), followed by addition of 4 M HCl in dioxane (1.2 mL). Afterdeprotection was complete as confirmed by LCMS, the solution wasconcentrated by rotary evaporation and dried under high vacuum overnightto yield the title compound (11 mg, 0.021 mmol, 0.4% yield). LCMS[M−H]⁻: 494.5. ¹H NMR (400 MHz, DMSO-d₆) δ 6.83-6.75 (m, 2H), 6.64-6.53(m, 1H), 5.58 (d, J=3.6 Hz, 1H), 5.53 (t, J=9.9 Hz, 1H), 5.03-4.90 (m,2H), 3.89 (t, J=10.9 Hz, 1H), 3.73 (dd, J=10.9, 5.9 Hz, 1H), 2.38-2.12(m, 6H), 1.58-1.39 (m, 6H), 0.92-0.76 (m, 9H).

Compound 45: 3,4,5-tris(butyryloxy)benzoic acid

Gallic Acid (400 mg, 2.35 mmol) was dissolved in pyridine (15 eq, 2.83mL, 35.2 mmol) in a dry round bottom flask. The flask was flushed withN₂ and the solution was chilled to 0° C. in an ice bath. Butyricanhydride (6 eq, 2.30 mL, 14.1 mmol) was added dropwise under N₂. Theresulting stirred solution was allowed to come to room temperature andreaction was monitored to completion by LCMS. The solution was dilutedwith 20 mL of ethyl acetate and washed with 1M HCl (20 mL) and saturatedNaCl (20 mL). The organic layer was dried over magnesium sulfate,filtered, and concentrated by rotary evaporation. The crude residue waspurified by flash chromatography (C18, 10-90% acetonitrile in water) andfractions were concentrated and fully dried by lyophilization to yieldcompound 45 (740 mg, 82.8% yield) as a white solid. LCMS [M−H]⁻: 379.1.¹H NMR (400 MHz, DMSO-d₆) δ 13.45 (bs, 1H), 7.74 (s, 2H), 2.62-2.55 (m,6H), 1.70-1.58 (m, 6H), 0.99-0.93 (m, 9H).

Compound 46:(2S,3S,4S,5R,6R)-3,4,5,6-tetrakis(propanoyloxy)oxane-2-carboxylic acid

Compound 47:(2S,3S,4S,5R,6S)-3,4,5,6-tetrakis(propanoyloxy)oxane-2-carboxylic acid

Step 1

To a solution of(2S,3S,4S,5R)-3,4,5,6-tetrahydroxytetrahydropyran-2-carboxylic acid (5g, 25.75 mmol, 1 eq) in propionic anhydride (25 mL) was added I2 (653.68mg, 2.58 mmol, 518.79 uL, 0.1 eq). The mixture was stirred at 25° C. for12 hr. TLC indicated(2S,3S,4S,5R)-3,4,5,6-tetrahydroxytetrahydropyran-2-carboxylic acid wasconsumed completely. The reaction mixture was concentrated under reducedpressure. Then the residue was taken up in toluene followed bydistillation in vacuum. The crude product propionic(2S,3S,4S,5R)-3,4,5,6-tetrakis(propionyloxy)tetrahydro-2H-pyran-2-carboxylic anhydride (7 g, crude) was obtained asa brown liquid.

Step 2

To a solution of propionic(2S,3S,4S,5R)-3,4,5,6-tetrakis(propionyloxy)tetrahydro-2H-pyran-2-carboxylicanhydride (7 g, 16.73 mmol, 1 eq) in DCM (70 mL) was added BnOH (3.62 g,33.46 mmol, 3.48 mL, 2 eq). The mixture was stirred at 25° C. for 12 hr.TLC indicated (propionic(2S,3S,4S,5R)-3,4,5,6-tetrakis(propionyloxy)tetrahydro-2H-pyran-2-carboxylicanhydride was consumed completely and one major new spot was detected.The reaction mixture was concentrated under reduced pressure. Theresidue was purified by column chromatography (SiO₂, Petroleumether/Ethyl acetate=20/1 to 6:1). Compound benzyl(2S,3S,4S,5R)-3,4,5,6-tetra(propanoyloxy)tetrahydropyran-2-carboxylate(1.5 g, crude) was obtained as a yellow oil. The residue was purified byprep-HPLC ([water (10 mM NH₄HCO3)-ACN]). The compound 3 for benzyl(2S,3S,4S,5R,6R)-3,4,5,6-tetra(propanoyloxy)tetrahydropyran-2-carboxylate (30 mg) was obtained as a white solid. Thecompound 3A benzyl(2S,3S,4S,5R,6S)-3,4,5,6-tetra(propanoyloxy)tetrahydropyran-2-carboxylate(100 mg) was obtained as a white solid. The compound ID was temporaryassigned.

Step 3

To a solution of benzyl(2S,3S,4S,5R,6R)-3,4,5,6-tetra(propanoyloxy)tetrahydropyran-2-carboxylate(30 mg, 59.00 umol, 1 eq) in THF (5 mL) was added Pd/C (3 mg, 59.00umol, 10% purity, 1.00 eq). The suspension was degassed and purged withH₂ for 3 times. The mixture was stirred under H₂ (15 Psi) at 25° C. for12 hr. LC-MS showed the desired compound was detected. Filtered andconcentrated under reduced pressure to give a residue. The residue waspurified by prep-HPLC ([water (0.1% TFA)-ACN]). The compound 46 for(2S,3S,4S,5R,6R)-3,4,5,6-tetra (propanoyloxy)tetrahydropyran-2-carboxylic acid (5.3 mg, 12.67 umol, 21.47% yield,100% purity) was obtained as a yellow solid. The compound ID wastemporary assigned. The structure was not further confirmed by 2D NMR.LCMS: (M+18)+: 436.1. @ 3.215 min. ¹H NMR (400 MHz, CDCl₃): δ 6.35 (s,1H), 5.49 (t, J=9.6 Hz, 1H), 5.22 (t, J=9.8 Hz, 1H), 5.07 (d, J=10.1 Hz,1H), 4.42 (s, 1H), 2.50-2.36 (m, 2H), 2.34-2.14 (m, 6H), 1.13 (t, J=7.6Hz, 3H), 1.07-0.97 (m, 9H)

Step 4

To a solution of benzyl(2S,3S,4S,5R,6S)-3,4,5,6-tetra(propanoyloxy)tetrahydropyran-2-carboxylate(50.00 mg, 98.33 umol, 1 eq) in THF (5 mL) was added Pd/C (3 mg, 98.33umol, 10% purity, 1.00 eq). The suspension was degassed and purged withH₂ for 3 times. The mixture was stirred under H₂ (15 Psi) at 25° C. for12 hr. LC-MS showed the desired compound was detected. The reactionmixture was filtered and concentrated under reduced pressure. Theresidue was purified by prep-HPLC ([water (0.1% TFA)-ACN]). The compound47 for(2S,3S,4S,5R,6R)-3,4,5,6-tetra(propanoyloxy)tetrahydropyran-2-carboxylicacid (11 mg, 26.29 umol, 26.74% yield, 100% purity) was obtained asyellow oil. The compound ID was temporary assigned. The structure wasnot further confirmed by 2D NMR. LCMS: (M+18)+: 436.1 @ 3.125 min. ¹HNMR (400 MHz, CDCl₃): δ 5.84 (d, J=7.6 Hz, 1H), 5.42-5.27 (m, 2H), 5.19(t, J=8.3 Hz, 1H), 4.28 (d, J=9.2 Hz, 1H), 2.46-2.24 (m, 8H), 1.18-1.06(m, 12H)

Compound 48:(2S,3S,4S,5R,6R)-3,4,5,6-tetrakis(butanoyloxy)oxane-2-carboxylic acid

This compound was prepared according to a modified procedure describedfor the preparation of compounds 46 and 47. LCMS: (M+Na⁺): 492.2. ¹H-NMR(400 MHz, CDCl₃): δ 6.26 (d, J=3.7 Hz, 1H), 5.49 (t, J=9.9 Hz, 1H), 5.16(t, J=10.0 Hz, 1H), 5.05 (dd, J=10.2, 3.7 Hz, 1H), 4.12 (d, J=10.3 Hz,1H), 2.34 (t, J=7.4 Hz, 2H), 2.28-2.09 (m, 6H), 1.69-1.58 (m, 2H),1.58-1.42 (m, 6H), 0.95-0.78 (m, 12H)

Compound 49:(2S,3S,4S,5R,6S)-3,4,5,6-tetrakis({[3-(1H-indol-3-yl)propanoyl]oxy})oxane-2-carboxylic

To a mixture of(2S,3S,4S,5R)-3,4,5,6-tetrahydroxytetrahydropyran-2-carboxylic acid (0.2g, 1.03 pGp-135 C₃ mmol, 1 eq) and 3-(1H-indol-3-yl)propanoic acid (1.17g, 6.18 mmol, 6 eq) in DCM (10 mL) was added DIPEA (1.07 g, 8.24 mmol,1.44 mL, 8 eq) and COMU (2.65 g, 6.18 mmol, 6 eq) in one portion at 25°C. under N2. The mixture was stirred at 50° C. for 12 hours. LCMS showedthe desired mass was detected. The reaction mixture was filtered andconcentrated under reduced pressure to give a residue. The residue waspurified by prep-HPLC (column: Waters Xbridge Prep OBD C18 150*40 10 u;mobile phase: [water (10 mM NH₄HCO₃)-ACN]; B %: 30%-55%,11 min) to give(2S,3S,4S,5R)-3,4,5,6-tetrakis[3-(1H-indol-3-yl)propanoyloxy]tetrahydropyran-2-carboxylicacid (88 mg, 8.77 umol, 8.51 e-1% yield, 96.37% purity) as a whitesolid. LCMS: (M−H⁺) 877.2 @ 1.375 min. LCMS: (M+18) 896.3 @ 2.832 min.¹H NMR: (400 MHz, Methanol-d4): δ 7.5-6.8 (m, 20H), 5.8 (d, 1H), 5.4-5.3(m, 1H), 5.3-5.2 (m, 1H), 5.2-5.1 (m, 1H), 4.1 (d, 1H), 3.0-2.0 (m, 16H)

Compound 50: (2R,3R,4R,5R)-3,5-bis(butanoyloxy)-2-methoxyoxan-4-ylbutanoate

This compound was prepared according to a modified procedure describedfor the preparation of compound 219.

LCMS: (m+H+)=375.4. ¹H NMR (400 MHz, DMSO-d6) δ 5.28 (t, 1H), 5.08 (q,1H), 4.90 (td, 1H), 4.69 (d, 1H), 3.92 (dd, 1H), 3.68 (dd, 1H), 3.34 (s,3H), 2.39-2.16 (m, 6H), 1.54 (m, 6H), 0.88 (m, 9H).

Compound 51: (2R,3R,4R,5R)-2-methoxy-3,5-bis(propanoyloxy)oxan-4-ylpropanoate

This compound was prepared according to a modified procedure describedfor the preparation of compound 219.

LCMS: (m+H+)=355.3. ¹H NMR (400 MHz, DMSO-d6) δ 5.26 (t, 1H), 5.07 (q,1H), 4.94-4.84 (m, 1H), 4.71 (d, 1H), 3.92 (dd, 1H), 3.69 (dd, 1H), 3.35(s, 3H), 2.45-2.17 (m, 6H), 1.02 (dt, 9H)

Compound 52: (2S,3R,4S,5S)-3,5-bis(butanoyloxy)-2-methoxyoxan-4-ylbutanoate

This compound was prepared according to a modified procedure describedfor the preparation of compound 219.

LCMS: (M+Na+) 397.3. ¹H NMR (400 MHz, DMSO-d6) δ 5.27 (dt, 1H), 5.24(dd, 1H), 5.04 (dd, 1H), 4.90 (d, 1H), 3.89 (dd, 1H), 3.61 (dd, 1H),3.32 (s, 3H), 2.39-2.13 (m, 6H), 1.65-1.42 (m, 6H), 0.97-0.80 (m, 9H).

Compound 53: (2S,3R,4S,5S)-2-methoxy-3,5-bis(propanoyloxy)oxan-4-ylpropanoate

This compound was prepared according to a modified procedure describedfor the preparation of compound 219.

LCMS: (M+Na+) 355.3. ¹H NMR (400 MHz, DMSO-d6) δ 5.27 (dt, 1H), 5.23(dd, 1H), 5.04 (dd, 1H), 4.90 (d, 1H), 3.89 (dd, 1H), 3.62 (dd, 1H),3.32 (s, 3H), 2.45-2.10 (m, 6H), 1.12-0.91 (m, 9H).

Compound 54:[(2R,3R,4S,5R,6S)-3,4,5-tris(butanoyloxy)-6-methoxyoxan-2-yl]methylbutanoate

This compound was prepared according to a modified procedure describedfor the preparation of compound 219.

LCMS: (M+Na+) 497.2. ¹H NMR (400 MHz, DMSO-d₆) δ 5.34 (dd, 1H), 5.00 (t,1H), 4.92 (d, 1H), 4.83 (dd, 1H), 4.14 (dd, 1H), 4.10-3.88 (m, 2H), 3.34(s, 3H), 2.35-2.08 (m, 8H), 1.60-1.40 (m, 8H), 0.93-0.78 (m, 12H)

Compound 55:[(2R,3R,4S,5R,6S)-6-methoxy-3,4,5-tris(propanoyloxy)oxan-2-yl]methylpropanoate

This compound was prepared according to a modified procedure describedfor the preparation of compound 219.

LCMS: (M+Na+) 441.2. ¹H NMR (400 MHz, DMSO-d6) δ 5.32 (dd, 1H), 4.99 (t,1H), 4.92 (d, 1H), 4.85 (dd, 1H), 4.17 (dd, 1H), 4.06 (dd, 1H), 3.93(ddd, 1H), 3.34 (s, 3H), 2.39-2.11 (m, 8H), 1.09-0.92 (m, 12H)

Compound 56:[(2R,3S,4S,5R,6S)-3,4,5-tris(butanoyloxy)-6-methoxyoxan-2-yl]methylbutanoate

This compound was prepared according to a modified procedure describedfor the preparation of compound 219.

LCMS: (M+Na+) 497.2. ¹H NMR (400 MHz, DMSO-d₆) δ 5.40 (dd, 1H), 5.27(dd, 1H), 5.04 (dd, 1H), 4.97 (d, 1H), 4.22 (t, 1H), 4.15-3.97 (m, 2H),3.35 (s, 3H), 2.47-2.07 (m, 8H), 1.69-1.38 (m, 8H), 1.01-0.77 (m, 12H)

Compound 57:[(2R,3S,4S,5R,6S)-6-methoxy-3,4,5-tris(propanoyloxy)oxan-2-yl]methylpropanoate

This compound was prepared according to a modified procedure describedfor the preparation of compound 219.

LCMS: (M+Na+): 441.1. ¹H NMR (400 MHz, DMSO-d₆) δ 5.40 (dd, 1H), 5.27(dd, 1H), 5.05 (dd, 1H), 4.98 (d, 1H), 4.22 (ddd, 1H), 4.07 (d, 2H),3.36 (s, 3H), 2.49-2.11 (m, 8H), 1.15-0.94 (m, 12H)

Compound 58:[(2R,3R,4S,5R,6R)-3,4,5-tris(butanoyloxy)-6-methoxyoxan-2-yl]methylbutanoate

This compound was prepared according to a modified procedure describedfor the preparation of compound 219.

LCMS: (M+Na+): 497.1. ¹H NMR (400 MHz, DMSO-d₆) δ 5.33 (t, 1H), 4.96 (t,1H), 4.81 (dd, 1H), 4.19 (dd, 1H), 4.12-3.97 (m, 2H), 3.39 (s, 3H),2.38-2.10 (m, 8H), 1.64-1.38 (m, 8H), 0.97-0.77 (m, 12H)

Compound 59:[(2R,3R,4S,5R,6R)-6-methoxy-3,4,5-tris(propanoyloxy)oxan-2-yl]methylpropanoate

This compound was prepared according to a modified procedure describedfor the preparation of compound 219.

LCMS: (M+Na+) 441.1. ¹H NMR (400 MHz, DMSO-d₆) δ 5.31 (t, 1H), 4.95 (t,1H), 4.80 (dd, 1H), 4.74 (d, 1H), 4.24 (dd, 1H), 4.10-3.99 (m, 2H), 3.39(s, 3H), 2.41-2.13 (m, 8H), 1.09-0.91 (m, 12H)

Compound 60:[(2R,3S,4S,5R,6R)-3,4,5-tris(butanoyloxy)-6-methoxyoxan-2-yl]methylbutanoate

This compound was prepared according to a modified procedure describedfor the preparation of compound 219.

LCMS: (M+Na+) 497.1. ¹H NMR (400 MHz, DMSO-d6) δ 5.30 (dd, 1H), 5.22(dd, 1H), 4.99 (dd, 1H), 4.64 (d, 1H), 4.28-4.20 (m, 1H), 4.15-3.96 (m,2H), 3.38 (s, 3H), 2.43-2.06 (m, 8H), 1.53 (ddq, 8H), 0.99-0.79 (m,12H).

Compound 61:[(2R,3S,4S,5R,6R)-6-methoxy-3,4,5-tris(propanoyloxy)oxan-2-yl]methylpropanoate

This compound was prepared according to a modified procedure describedfor the preparation of compound 219.

LCMS (M+Na): 441.1. ¹H NMR (400 MHz, DMSO-d6) δ 5.30 (dd, 1H), 5.20 (dd,1H), 4.98 (dd, 1H), 4.65 (d, 1H), 4.24 (td, 1H), 4.18-4.01 (m, 2H), 3.39(s, 3H), 2.48-2.08 (m, 8H), 1.03 (ddt, 12H)

Compound 62:(2S,3R,4R,5S)-6-hydroxy-4,5-bis({[3-(1H-indol-3-yl)propanoyl]oxy})-2-methyloxan-3-yl3-(1H-indol-3-yl)propanoate

This compound was prepared according to a procedure described forcompound 37 with the exception that a starting material for compound 62was used and that the synthesis was stopped at the stage when compound62 was produced. LCMS: (M−H): 676.3. ¹H NMR (400 MHz, Chloroform-d) δ7.77-6.47 (m, 18H), 5.42-5.32 (m, 1H), 5.29-5.22 (m, 1H), 5.12-5.03 (m,1H), 4.69-4.62 (m, 1H), 4.41-4.35 (m, 1H), 3.87-3.81 (m, 1H), 3.20-3.00(m, 4H), 2.90-2.75 (m, 4H), 2.74-2.59 (m, 2H), 2.12-1.97 (m, 2H),1.22-1.07 (m, 3H)

Compound 63:(2S,3S,4S,5R,6S)-3,4,5,6-tetrakis({[2-(1H-indol-3-yl)acetyl]oxy})oxane-2-carboxylicacid

To a mixture of(2S,3S,4S,5R,6S)-3,4,5,6-tetrahydroxytetrahydropyran-2-carboxylic acid(200 mg, 1.03 mmol, 1 eq) and 2-(1H-indol-3-yl)acetic acid (1.08 g, 6.18mmol, 6 eq) in DCM (10 mL) was added COMU (2.65 g, 6.18 mmol, 6 eq) andDIPEA (1.07 g, 8.24 mmol, 1.44 mL, 8 eq) in one portion at 25° C. underN₂. The mixture was stirred at 50° C. for 12 hours. LCMS showed thedesired mass was detected. The reaction mixture was filtered andconcentrated under reduced pressure to give a residue. The residue waspurified by prep-HPLC (column: Waters Xbridge Prep OBD C18 150*40 10 u;mobile phase: [water (10 mM NH₄HCO₃)-MeOH]; B %: 25%-45%,11 min) to givecompound 63(2S,3S,4S,5R)-3,4,5,6-tetrakis(2-(1H-indol-3-yl)acetoxy)tetrahydro-2H-pyran-2-carboxylicacid (5 mg, 5.48 μmol, 0.532% yield, 90.23% purity) as a light yellowsolid. LCMS: (M+18)⁺840.2 @ 2.594 min. ¹H NMR (400 MHz, Methanol-d₄) δ7.53-6.79 (m, 20H), 5.69 (d, J=8.3 Hz, 1H), 5.39-5.02 (m, 3H), 4.61 (s,1H), 3.67-3.18 (m, 4H), 3.15-2.90 (m, 4H).

Compound 64:(2S,3S,4R,5R,6S)-3,4,5-tris({[3-(1H-indol-3-yl)propanoyl]oxy})-6-methyloxan-2-yl3-(1H-indol-3-yl)propanoate

This compound was prepared following a modified procedure described forcompound 63. ¹H NMR (400 MHz, Chloroform-d) δ 7.88-6.20 (m, 24H),5.45-5.20 (m, 3H), 5.15-4.88 (m, 1H), 4.16-3.89 (m, 1H), 3.24-2.99 (m,4H), 2.99-2.65 (m, 8H), 2.41-1.99 (m, 4H), 1.37-0.91 (m, 3H).

Compound 65:[(2R,3R,4S,5S,6S)-3,4,5-tris(butanoyloxy)-6-methoxyoxan-2-yl]methylbutanoate

This compound was prepared according to a modified procedure describedfor the preparation of compound 219.

LCMS: (M+Na+) 497.2. ¹H NMR (400 MHz, DMSO-d6) δ 5.28-5.08 (m, 3H), 4.78(d, 1H), 4.21-4.07 (m, 2H), 3.94 (ddd, 1H), 3.36 (s, 3H), 2.44-2.11 (m,8H), 1.69-1.40 (m, 8H), 0.89 (m, 12H)

Compound 66:[(2R,3R,4S,5S,6S)-6-methoxy-3,4,5-tris(propanoyloxy)oxan-2-yl]methylpropanoate

This compound was prepared according to a modified procedure describedfor the preparation of compound 219.

LCMS: (M+Na+) 441.1. ¹H NMR (400 MHz, DMSO-d6) δ 5.25-5.15 (m, 1H), 5.14(m, 2H), 4.80 (d, 1H), 4.20 (dd, 1H), 4.10 (dd, 1H), 3.95 (m, 1H), 3.37(s, 3H), 2.48-2.18 (m, 8H), 1.14-0.93 (m, 12H)

Compound 67: (2R,3S,4R,5R)-4,5-bis(butanoyloxy)-2-methoxyoxan-3-ylbutanoate

This compound was prepared according to a modified procedure describedfor the preparation of compound 219.

LCMS: (M+Na+) 397.2. ¹H NMR (400 MHz, DMSO-d6) δ 5.31-5.20 (m, 2H), 5.04(dd, 1H), 4.90 (d, 1H), 3.89 (dd, 1H), 3.62 (dd, 1H), 3.32 (s, 3H),2.40-2.13 (m, 6H), 1.65-1.42 (m, 6H), 0.97-0.78 (m, 9H)

Compound 68: (2R,3S,4R,5R)-2-methoxy-3,5-bis(propanoyloxy)oxan-4-ylpropanoate

This compound was prepared according to a modified procedure describedfor the preparation of compound 219.

LCMS: (M+Na+) 355.1. ¹H NMR (400 MHz, DMSO-d6) δ 5.31-5.22 (m, 2H), 5.06(dd, 1H), 4.92 (d, 1H), 3.91 (dd, 1H), 3.64 (dd, 1H), 3.34 (s, 3H),2.47-2.12 (m, 6H), 1.13-0.95 (m, 9H)

Compound 69:(2S,3R,4S,5S)-3,4,5-tris({[3-(1H-indol-3-yl)propanoyl]oxy})oxan-2-yl3-(1H-indol-3-yl)propanoate

This compound was prepared following a modified procedure described forcompound 63.

LCMS: (M+H⁺):835.3. ¹H NMR (400 MHz, Chloroform-d) δ 7.84 (d, J=6.9 Hz,2H), 7.69 (s, 1H), 7.59-7.45 (m, 5H), 7.34-7.25 (m, 2H), 7.22-7.06 (m,10H), 7.03 (d, J=2.4 Hz, 1H), 6.92 (d, 0=2.3 Hz, 1H), 6.76 (d, J=2.3 Hz,1H), 6.49 (d, J=2.3 Hz, 1H), 5.68 (d, J=6.9 Hz, 1H), 5.37-5.28 (m, 2H),5.07 (dd, J=9.2, 3.4 Hz, 1H), 3.95 (dd, J=13.0, 3.7 Hz, 1H), 3.74 (dd,J=13.0, 2.0 Hz, 1H), 3.20-2.57 (m, 12H), 2.55-2.39 (m, 2H), 2.24-1.98(m, 2H).

Compound 70:(2R,3S,4R,5R,6S)-3,4,5-tris({[3-(1H-indol-3-yl)propanoyl]oxy})-6-methyloxan-2-yl3-(1H-indol-3-yl)propanoate

This compound was prepared following a modified procedure described forcompound 63. ¹H NMR (400 MHz, Chloroform-d) δ 7.78 (s, 1H), 7.74 (s,1H), 7.66 (s, 1H), 7.54-7.46 (m, 2H), 7.51-7.37 (m, 2H), 7.29 (s, 1H),7.26-7.16 (m, 2H), 7.16-6.92 (m, 11H), 6.87 (d, J=2.3 Hz, 1H), 6.61 (d,J=2.3 Hz, 1H), 6.30 (d, J=2.4 Hz, 1H), 5.67 (d, J=8.3 Hz, 1H), 5.36 (dd,J=10.5, 8.3 Hz, 1H), 5.27-5.21 (m, 1H), 4.98 (dd, J=10.5, 3.3 Hz, 1H),3.98-3.86 (m, 1H), 3.18-2.51 (m, 12H), 2.40-2.22 (m, 2H), 1.99-1.76 (m,2H), 1.21-1.11 (m, 3H)

Compound 71: (2S,3R,4S,5R)-4,5-bis(butanoyloxy)-2-methoxyoxan-3-ylbutanoate

This compound was prepared according to a modified procedure describedfor the preparation of compound 219.

LCMS: (M+Na+) 397.2. ¹H NMR (400 MHz, DMSO-d₆) δ 5.37 (t, 1H), 4.97(ddd, 1H), 4.91 (d, 1H), 4.86 (dd, 1H), 3.80 (dd, 1H), 3.50 (t, 1H),3.36 (s, 3H), 2.37-2.13 (m, 6H), 1.53 (qd, 6H), 0.89 (td, 9H)

Compound 72: (2S,3R,4S,5R)-2-methoxy-4,5-bis(propanoyloxy)oxan-3-ylpropanoate

This compound was prepared according to a modified procedure describedfor the preparation of compound 219.

LCMS: (M+Na+) 355.1. ¹H NMR (400 MHz, DMSO-d₆) δ 5.35 (t, 1H), 4.96(ddd, 1H), 4.91 (d, 1H), 4.87 (dd, 1H), 3.80 (dd, 1H), 3.51 (t, 1H),3.36 (s, 3H), 2.37-2.23 (m, 6H), 1.02 (td, 9H)

Compound 73: (2R,3R,4S,5R)-2-methoxy-4,5-bis(propanoyloxy)oxan-3-ylpropanoate

This compound was prepared according to a modified procedure describedfor the preparation of compound 219.

LCMS: (M+Na+) 355.1. ¹H NMR (400 MHz, DMSO-d6) δ 5.24 (t, 1H), 4.90 (td,1H), 4.81 (dd, 1H), 4.64 (d, 1H), 4.01 (dd, 1H), 3.55 (dd, 1H), 3.40 (s,3H), 2.38-2.22 (m, 6H), 1.08-0.96 (m, 9H).

Compound 74: (2S,3R,4S,5S)-3,4,5-tris(propanoyloxy)oxan-2-yl3-(1H-indol-3-yl)propanoate

Step 1

Propionic anhydride (500 mL, 4 mol, 10 eq) was added to L-arabinose (60g, 0.4 mol, 1 eq) in a 2 L round bottom flask equipped with a stirbar.Pyridine (320 mL, 4 mol, 10 eq) was added to the flask, and the reactionwas stirred overnight at room temperature. The reaction was washed with1M HCl, saturated sodium bicarbonate, and brine. Next the propionicanhydride was removed by rotary evaporation to yield 170 g of crude(2S,3R,4S,5S)-tetrahydro-2H-pyran-2,3,4,5-tetrayl tetrapropionate.Product was taken forward to next step without further purification.

Step 2

Benzylamine (78.5 mL, 720 mmol, 5 equiv) was added to a stirred solutionof (2S,3R,4S,5S)-tetrahydro-2H-pyran-2,3,4,5-tetrayl tetrapropionate (62g, 144 mmol, 1 equiv) in THF (500 mL) at RT. When the TLC indicatedcomplete disappearance of starting material (4-8 h), the reaction wasquenched by addition of 1M HCl (375 mL), and the mixture was extractedwith ethyl acetate (3×500 mL). The organic phase was dried, pulledthrough a plug of silica, and concentrated. The crude product waspurified by using column chromatography (100% hexanes to 50% Ethylacetate in hexanes) to yield(3R,4S,5S)-2-hydroxytetrahydro-2H-pyran-3,4,5-triyl tripropionate (16 g,44.3 mmol, 30.8% yield).

Step 3

Indole-propionic acid (23.0 g, 122 mmol, 1.5 eq), EDC HCl (23.4 g, 122mmol, 1.5 eq), and DMAP (15 g, 122 mmol, 1.5 eq) were stirred in DCM(200 mL) at room temperature for a few minutes. Compound(3R,4S,5S)-2-hydroxytetrahydro-2H-pyran-3,4,5-triyl tripropionate (26 g,81.6 mmol, 1 eq) was added and the solution was stirred overnight. Thesolution was washed with saturated ammonium chloride, saturated sodiumbicarbonate, and brine, then loaded onto silica and purified by columnchromatography (100% hexanes to 50% Ethyl acetate in hexanes) to yieldthe title compound (10.7 g, 21.8 mmol, 26.8% yield) as a gooey solid.LCMS (M+Na⁺): 512.2. ¹H NMR (400 MHz, DMSO-d₆) δ 10.78 (s, 1H), 7.48(dd, J=7.7, 1.0 Hz, 1H), 7.31 (dd, J=8.1, 1.1 Hz, 1H), 7.10-7.01 (m,2H), 7.00-6.91 (m, 1H), 5.77 (d, J=7.6 Hz, 1H), 5.28 (dd, J=9.7, 3.6 Hz,1H), 5.22-5.16 (m, 1H), 5.09 (dd, J=9.7, 7.6 Hz, 1H), 3.98 (dd, J=13.2,1.7 Hz, 1H), 3.87 (dd, J=13.0, 2.8 Hz, 1H), 2.92 (t, J=7.4 Hz, 2H), 2.70(td, J=7.8, 7.4, 2.9 Hz, 2H), 2.37 (q, J=7.5 Hz, 2H), 2.27-2.05 (m, 4H),1.04 (t, J=7.5 Hz, 3H), 0.95 (t, J=7.5 Hz, 3H), 0.89 (t, J=7.5 Hz, 3H).

Compound 75: (2R,3R,4S,5R)-3,4,5-tris(propanoyloxy)oxan-2-yl3-(1H-indol-3-yl)propanoate

This compound was prepared following a modified procedure described forcompound 74. ¹H NMR (400 MHz, DMSO-d6) δ 10.79 (s, 1H), 7.56-7.43 (m,1H), 7.31 (m, 1H), 7.13 (d, 1H), 7.05 (ddd, 1H), 6.96 (ddd, 1H), 6.13(d, 1H), 5.33 (t, 1H), 5.06-4.92 (m, 2H), 3.80 (dd, 1H), 3.52 (t, 1H),3.05-2.79 (m, 4H), 2.32-2.03 (m, 6H), 0.97 (m, 6H), 0.89 (t, 3H)

Compound 76:(2S,3R,4S,5R)-2-{[2-(1H-indol-3-yl)acetyl]oxy}-4,5-bis(propanoyloxy)oxan-3-ylpropanoate

This compound was prepared following a modified procedure described forcompound 74.

LCMS: (M+Na+):498.2. ¹H NMR (400 MHz, Chloroform-d) δ 8.07 (s, 1H),7.60-7.53 (m, 1H), 7.35 (dt, J=8.2, 1.0 Hz, 1H), 7.24-7.16 (m, 2H),7.16-7.09 (m, 1H), 5.74 (d, J=7.1 Hz, 1H), 5.19 (t, J=8.4 Hz, 1H), 5.05(dd, J=8.6, 7.0 Hz, 1H), 5.02-4.93 (m, 1H), 4.11 (dd, J=12.0, 5.0 Hz,1H), 3.81 (d, J=0.9 Hz, 2H), 3.49 (dd, J=12.0, 8.7 Hz, 1H), 2.36-2.17(m, 4H), 2.12-1.85 (m, 2H), 1.14-1.03 (m, 6H), 0.94 (t, J=7.6 Hz, 3H).

Compound 77: (2R,3S,4R,5R,6S)-6-methyl-3,4,5-tris(propanoyloxy)oxan-2-yl3-(1H-indol-3-yl)propanoate

This compound was prepared following a modified procedure described forcompound 74. LCMS: (M+Na+):526.2. ¹H NMR (400 MHz, Chloroform-d) δ 7.97(s, 1H), 7.57-7.47 (m, 1H), 7.32-7.23 (m, 1H), 7.11 (ddd, J=8.1, 7.1,1.3 Hz, 1H), 7.04 (ddd, J=8.0, 7.0, 1.1 Hz, 1H), 6.97-6.89 (m, 1H), 5.66(d, J=8.4 Hz, 1H), 5.28 (dd, J=10.4, 8.3 Hz, 1H), 5.21 (dd, J=3.5, 1.1Hz, 1H), 5.03 (dd, J=10.4, 3.4 Hz, 1H), 3.89 (qd, J=6.4, 1.2 Hz, 1H),3.13-2.95 (m, 2H), 2.79-2.61 (m, 2H), 2.49-2.30 (m, 2H), 2.21-1.93 (m,4H), 1.26-1.09 (m, 6H), 1.00 (t, J=7.5 Hz, 3H), 0.91 (t, J=7.6 Hz, 3H)

Compound 78: (2S,3R,4R,5S,6S)-6-methyl-3,4,5-tris(propanoyloxy)oxan-2-ylpropanoate

This compound was prepared according to a modified procedure describedfor the preparation of compound 219.

LCMS: (M+Na⁺):411.1. ¹H NMR (400 MHz, Chloroform-cQ δ 6.03 (d, J=1.9 Hz,1H), 5.33 (dd, J=10.1, 3.5 Hz, 1H), 5.27 (dd, J=3.5, 2.0 Hz, 1H), 5.15(t, J=10.0 Hz, 1H), 4.00-3.88 (m, 1H), 2.51-2.37 (m, 4H), 2.37-2.19 (m,4H), 1.27-1.04 (m, 15H).

Compound 79:(2R,3R,4S,5R)-4,5-bis(butanoyloxy)-2-{[(2R,3R,4S,5R)-3,4,5-tris(butanoyloxy)oxan-2-yl]oxy}oxan-3-ylbutanoate

This compound was prepared according to a modified procedure describedfor the preparation of compound 219.

Compound 80:(2S,3R,4R,5R)-3,4,5-tris({[3-(1H-indol-3-yl)propanoyl]oxy})oxan-2-yl3-(1H-indol-3-yl)propanoate

This compound was prepared following a modified procedure described forcompound 63. LCMS: (M+H⁺): 835.3. ¹H NMR (400 MHz, Chloroform-d) δ 7.77(s, 1H), 7.73 (s, 1H), 7.49 (d, J=8.1 Hz, 3H), 7.46-7.38 (m, 2H), 7.25(t, J=8.4 Hz, 3H), 7.19-6.92 (m, 10H), 6.83 (d, J=2.3 Hz, 1H), 6.77 (d,J=2.3 Hz, 1H), 6.75-6.67 (m, 1H), 6.65-6.60 (m, 1H), 6.03 (d, J=3.7 Hz,1H), 5.52 (s, 1H), 5.08 (t, J=3.5 Hz, 1H), 5.03-4.94 (m, 1H), 3.76 (t,J=10.4 Hz, 1H), 3.51 (dd, J=11.2, 4.7 Hz, 1H), 3.06-2.86 (m, 8H),2.67-2.38 (m, 8H).

Compound 81:(2R,3R,4S,5S)-4,5-bis(butanoyloxy)-2-{[3-(1H-indol-3-yl)propanoyl]oxy}oxan-3-ylbutanoate

This compound was prepared following a modified procedure described forcompound 74. LCMS: (M+H⁺): 532.2. ¹H NMR (400 MHz, Methanol-d₄) δ 7.56(d, J=7.8 Hz, 1H), 7.32 (d, J=8.1 Hz, 1H), 7.12-7.05 (m, 2H), 7.05-6.97(m, 1H), 6.26 (d, J=3.6 Hz, 1H), 5.36-5.26 (m, 2H), 5.21 (dd, J=10.5,3.6 Hz, 1H), 3.80 (dd, J=13.4, 1.3 Hz, 1H), 3.62 (dd, J=13.4, 1.9 Hz,1H), 3.21-3.04 (m, 2H), 2.87 (t, J=6.9 Hz, 2H), 2.44-2.34 (m, 2H), 2.22(t, J=7.2 Hz, 2H), 2.06-1.95 (m, 2H), 1.73-1.61 (m, 2H), 1.64-1.52 (m,2H), 1.45 (h, J=7.3 Hz, 2H), 0.98 (t, J=7.4 Hz, 3H), 0.91 (t, J=7.4 Hz,3H), 0.80 (t, J=7.4 Hz, 3H).

Compound 82:(2S,3R,4S,5S)-4,5-bis(butanoyloxy)-2-{[3-(1H-indol-3-yl)propanoyl]oxy}oxan-3-ylbutanoate

This compound was prepared following a modified procedure described forcompound 74. LCMS: (M+H⁺): 532.2. ¹H NMR (400 MHz, Methanol-d₄) δ 7.41(dt, J=7.9, 1.1 Hz, 1H), 7.21 (dt, J=8.2, 1.0 Hz, 1H), 7.02-6.94 (m,1H), 6.93-6.85 (m, 2H), 5.63 (d, J=7.2 Hz, 1H), 5.24-5.17 (m, 1H),5.17-5.05 (m, 2H), 3.83 (dd, J=13.1, 3.3 Hz, 1H), 3.76 (dd, J=13.1, 1.9Hz, 1H), 3.03-2.89 (m, 2H), 2.73-2.58 (m, 2H), 2.34-1.90 (m, 6H),1.64-1.53 (m, 2H), 1.52-1.43 (m, 2H), 1.41-1.31 (m, 2H), 0.94-0.84 (m,3H), 0.84-0.76 (m, 3H), 0.73 (t, J=7.4 Hz, 3H).

Compound 83:(2S,3R,4R,5S,6S)-4,5-bis(butanoyloxy)-2-{[3-(1H-indol-3-yl)propanoyl]oxy}-6-methyloxan-3-ylbutanoate

This compound was prepared following a modified procedure described forcompound 74.

Compound 84:(2S,3S,4R,5R,6S)-4,5-bis(butanoyloxy)-2-{[3-(1H-indol-3-yl)propanoyl]oxy}-6-methyloxan-3-ylbutanoate

This compound was prepared following a modified procedure described forcompound 74. ¹H NMR (400 MHz, Chloroform-d) δ 8.05 (s, 1H), 7.61 (d,J=7.8 Hz, 1H), 7.39-7.32 (m, 1H), 7.19 (ddd, J=8.2, 7.0, 1.3 Hz, 1H),7.13 (td, J=7.5, 1.2 Hz, 1H), 7.08 (d, J=2.3 Hz, 1H), 6.33 (d, J=3.2 Hz,1H), 5.40-5.22 (m, 3H), 4.01-3.91 (m, 1H), 3.20-3.08 (m, 2H), 2.84 (t,J=7.3 Hz, 2H), 2.47-2.32 (m, 2H), 2.20 (td, J=7.3, 1.8 Hz, 2H), 2.04(td, J=7.3, 1.2 Hz, 2H), 1.76-1.41 (m, 6H), 1.04-0.87 (m, 9H), 0.83 (t,J=7.4 Hz, 3H)

Compound 85:[(2R,3R,4S,5R,6S)-3,4,5-tris(butanoyloxy)-6-{[3-(1H-indol-3-yl)propanoyl]oxy}oxan-2-yl]methylbutanoate

This compound was prepared following a modified procedure described forcompound 74. ¹H NMR (400 MHz, Chloroform-d) δ 7.93 (s, 1H), 7.49 (dd,J=7.8, 1.1 Hz, 1H), 7.28 (dt, J=8.1, 1.0 Hz, 1H), 7.12 (ddd, J=8.1, 7.0,1.3 Hz, 1H), 7.04 (ddd, J=8.1, 7.0, 1.1 Hz, 1H), 6.92 (d, J=2.4 Hz, 1H),5.68 (d, 0=8.2 Hz, 1H), 5.26-5.17 (m, 1H), 5.14-5.05 (m, 2H), 4.14 (qd,0=12.5, 3.4 Hz, 2H), 3.77 (ddd, J=10.0, 4.5, 2.4 Hz, 1H), 3.06-2.97 (m,2H), 2.78-2.61 (m, 2H), 2.30-2.22 (m, 2H), 2.24-2.09 (m, 4H), 2.03 (td,J=7.4, 2.7 Hz, 2H), 1.63-1.36 (m, 8H), 0.91-0.72 (m, 12H).

Compound 86:(2R,3R,4R,5S,6S)-4,5-bis(butanoyloxy)-2-{[3-(1H-indol-3-yl)propanoyl]oxy}-6-methyloxan-3-ylbutanoate

This compound was prepared following a modified procedure described forcompound 74.

Compound 87:(2R,3S,4R,5R,6S)-4,5-bis(butanoyloxy)-2-{[3-(1H-indol-3-yl)propanoyl]oxy}-6-methyloxan-3-ylbutanoate

This compound was prepared following a modified procedure described forcompound 74. ¹H NMR (400 MHz, Chloroform-d) δ 7.96 (s, 1H), 7.57 (d,J=7.9 Hz, 1H), 7.35 (d, 0=8.1 Hz, 1H), 7.23-7.15 (m, 1H), 7.15-7.07 (m,1H), 7.00 (d, 0=2.3 Hz, 1H), 5.73 (d, 0=8.4 Hz, 1H), 5.41-5.30 (m, 1H),5.32-5.27 (m, 1H), 5.10 (dd, 0=10.4, 3.4 Hz, 1H), 4.02-3.93 (m, 1H),3.15-3.00 (m, 2H), 2.87-2.68 (m, 2H), 2.51-2.34 (m, 2H), 2.27-2.16 (m,2H), 2.16-2.00 (m, 2H), 1.78-1.66 (m, 2H), 1.62-1.43 (m, 4H), 1.27-1.18(m, 3H), 0.99 (t, J=7.4 Hz, 3H), 0.89 (t, J=7.4 Hz, 3H), 0.83 (t, J=7.4Hz, 3H)

Compound 88: (2R,3R,4S,5R)-4,5-bis(butanoyloxy)-2-methoxyoxan-3-ylbutanoate

This compound was prepared following a modified procedure described forcompounds 46 and 47. LCMS: (M+Na+) 397.2. ¹H NMR (400 MHz, DMSO-d6) δ5.26 (t, 1H), 4.90 (td, 1H), 4.82 (dd, 1H), 4.63 (d, 1H), 4.00 (dd, 1H),3.54 (dd, 1H), 3.40 (s, 3H), 2.34-2.18 (m, 6H), 1.59-1.48 (m, 6H),0.97-0.81 (m, 9H)

Compound 90:[(2R,3R,4S,5R,6R)-3,4,5-tris(butanoyloxy)-6-{[3-(1H-indol-3-yl)propanoyl]oxy}oxan-2-yl]methylbutanoate

This compound was prepared following a modified procedure described forcompound 74. ¹H NMR (400 MHz, Chloroform-d) δ 8.04 (s, 1H), 7.54 (dd,J=7.8, 1.2 Hz, 1H), 7.29 (dt, J=8.2, 1.0 Hz, 1H), 7.12 (ddd, J=8.2, 7.0,1.3 Hz, 1H), 7.09-7.00 (m, 2H), 6.26 (d, J=3.7 Hz, 1H), 5.37 (t, J=9.9Hz, 1H), 5.05 (t, J=9.9 Hz, 1H), 4.97 (dd, J=10.3, 3.7 Hz, 1H), 3.97(dd, J=12.5, 3.9 Hz, 1H), 3.86 (dd, J=12.5, 2.3 Hz, 1H), 3.71 (ddd,J=10.3, 4.0, 2.2 Hz, 1H), 3.17-3.00 (m, 2H), 2.81 (t, J=7.2 Hz, 2H),2.23 (td, J=7.4, 1.7 Hz, 2H), 2.20-2.08 (m, 4H), 2.00-1.87 (m, 2H),1.62-1.46 (m, 6H), 1.47-1.34 (m, 2H), 0.91-0.80 (m, 9H), 0.74 (t, J=7.4Hz, 3H).

Compound 91: (2S,3R,4S,5R)-3,4,5-tris(propanoyloxy)oxan-2-yl(2E)-3-(1H-indol-3-yl)prop-2-enoate

This compound was prepared following a modified procedure described forcompound 74. ¹H NMR (400 MHz, Chloroform-d) δ 8.50 (s, 1H), 7.98 (d,J=15.9 Hz, 1H), 7.94-7.88 (m, 1H), 7.54 (d, J=2.8 Hz, 1H), 7.46-7.39 (m,1H), 7.35-7.24 (m, 2H), 6.41 (d, J=15.9 Hz, 1H), 5.87 (d, J=7.2 Hz, 1H),5.31 (t, J=8.6 Hz, 1H), 5.21 (dd, J=8.8, 7.2 Hz, 1H), 5.07 (td, J=8.7,5.2 Hz, 1H), 4.20 (dd, J=11.9, 5.2 Hz, 1H), 3.58 (dd, J=11.9, 9.0 Hz,1H), 2.38-2.25 (m, 6H), 1.22-1.04 (m, 9H).

Compound 92: (2S,3R,4R,5R)-3,4,5-tris(propanoyloxy)oxan-2-yl(2E)-3-(1H-indol-3-yl)prop-2-enoate

This compound was prepared following a modified procedure described forcompound 74. ¹H NMR (400 MHz, Chloroform-d) δ 8.58 (s, 1H), 7.99 (d,J=15.9 Hz, 1H), 7.95-7.89 (m, 1H), 7.55 (d, J=2.8 Hz, 1H), 7.48-7.38 (m,1H), 7.35-7.24 (m, 2H), 6.44 (d, J=15.9 Hz, 1H), 6.19 (d, J=4.8 Hz, 1H),5.62 (t, J=3.5 Hz, 1H), 5.27-5.18 (m, 2H), 4.11 (dd, J=12.3, 3.4 Hz,1H), 3.95 (dd, 0=12.4, 5.8 Hz, 1H), 2.39 (ddd, J=9.8, 4.8, 2.2 Hz, 6H),1.21-1.10 (m, 9H)

Compound 93: (2R,3R,4S,5R)-3,4,5-tris(propanoyloxy)oxan-2-yl(2E)-3-(1H-indol-3-yl)prop-2-enoate

This compound was prepared following a modified procedure described forcompound 74.

Compound 94: (2R,3R,4R,5R)-3,4,5-tris(propanoyloxy)oxan-2-yl3-(1H-indol-3-yl)propanoate

This compound was prepared following a modified procedure described forcompound 74.

Compound 95:(2S,3R,4S,5R,6R)-3,4,5-tris(propanoyloxy)-6-[(propanoyloxy)methyl]oxan-2-yl(2E)-3-(1H-indol-3-yl)prop-2-enoate

This compound was prepared following a modified procedure described forcompound 74. ¹H NMR (400 MHz, Chloroform-d) δ 8.48 (s, 1H), 7.91 (d,J=15.9 Hz, 1H), 7.87-7.79 (m, 1H), 7.47 (d, J=2.9 Hz, 1H), 7.41-7.32 (m,1H), 7.28-7.16 (m, 2H), 6.33 (d, J=16.0 Hz, 1H), 5.84 (d, J=7.8 Hz, 1H),5.32-5.19 (m, 2H), 5.15 (t, J=9.5 Hz, 1H), 4.27 (dd, J=12.5, 4.6 Hz,1H), 4.08 (dd, 12.5, 2.2 Hz, 1H), 3.86 (ddd, J=10.0, 4.6, 2.2 Hz, 1H),2.37-2.14 (m, 8H), 1.11-0.95 (m, 12H).

Compound 96:(2R,3R,4S,5R)-4,5-bis(butanoyloxy)-2-{[3-(1H-indol-3-yl)propanoyl]oxy}oxan-3-ylbutanoate

This compound was prepared following a modified procedure described forcompound 74. ¹H NMR (400 MHz, Methanol-d₄) δ 7.58 (dt, J=7.8, 1.1 Hz,1H), 7.34 (dt, J=8.1, 1.0 Hz, 1H), 7.15-7.07 (m, 2H), 7.03 (ddd, J=8.0,7.0, 1.1 Hz, 1H), 6.28 (d, J=3.5 Hz, 1H), 5.41-5.28 (m, 2H), 5.23 (dd,J=10.5, 3.6 Hz, 1H), 3.83 (dd, J=13.3, 1.4 Hz, 1H), 3.65 (dd, J=13.3,2.0 Hz, 1H), 3.23-3.05 (m, 2H), 2.89 (t, J=6.9 Hz, 2H), 2.49-2.31 (m,2H), 2.24 (t, J=7.2 Hz, 2H), 2.08-1.98 (m, 2H), 1.76-1.54 (m, 4H),1.54-1.41 (m, 2H), 1.06-0.89 (m, 6H), 0.83 (t, J=7.4 Hz, 3H).

Compound 97:(2S,3R,4S,5R)-4,5-bis(butanoyloxy)-2-{[3-(1H-indol-3-yl)propanoyl]oxy}oxan-3-ylbutanoate

This compound was prepared following a modified procedure described forcompound 74. ¹H NMR (400 MHz, Methanol-d₄) δ 7.56-7.47 (m, 1H),7.37-7.30 (m, 1H), 7.10 (ddd, J=8.1, 6.9, 1.3 Hz, 1H), 7.06-6.97 (m,2H), 5.79-5.70 (m, 1H), 5.37-5.29 (m, 1H), 5.29-5.17 (m, 2H), 3.95 (dd,J=13.1, 3.3 Hz, 1H), 3.88 (dd, J=13.1, 1.9 Hz, 1H), 3.15-3.00 (m, 2H),2.83-2.72 (m, 2H), 2.48-2.34 (m, 2H), 2.22 (t, J=7.3 Hz, 2H), 2.18-2.00(m, 2H), 1.76-1.65 (m, 2H), 1.68-1.52 (m, 2H), 1.55-1.42 (m, 2H),1.06-0.96 (m, 3H), 0.99-0.80 (m, 6H).

Compound 98: (2S,3R,4R,5R)-3,4,5-tris(propanoyloxy)oxan-2-yl3-(1H-indol-3-yl)propanoate

This compound was prepared following a modified procedure described forcompound 74.

Compound 99:(2R,3R,4S,5R,6R)-3,4,5-tris(propanoyloxy)-6-[(propanoyloxy)methyl]oxan-2-yl(2E)-3-(1H-indol-3-yl)prop-2-enoate

This compound was prepared following a modified procedure described forcompound 74. ¹H NMR (400 MHz, Chloroform-d) δ 8.59 (s, 1H), 8.09-7.99(m, 2H), 7.61 (d, J=2.8 Hz, 1H), 7.52-7.43 (m, 1H), 7.39-7.31 (m, 2H),6.58-6.49 (m, 2H), 5.68 (t, J=9.9 Hz, 1H), 5.29-5.18 (m, 2H), 4.33 (dd,J=12.4, 4.2 Hz, 1H), 4.26 (ddd, J=10.3, 4.2, 2.1 Hz, 1H), 4.15 (dd,J=12.3, 2.1 Hz, 1H), 2.46-2.25 (m, 8H), 1.20-1.07 (m, 12H).

Compound 100: (2S,3R,4S,5S)-3,4,5-tris(propanoyloxy)oxan-2-yl(2E)-3-(1H-indol-3-yl)prop-2-enoate

This compound was prepared following a modified procedure described forcompound 74.

Compound 101: (2R,3R,4R,5R)-3,4,5-tris(propanoyloxy)oxan-2-yl propanoate

This compound was prepared according to a modified procedure describedfor the preparation of compound 219.

Compound 102:(2S,3R,4R,5S,6S)-6-methyl-3,4,5-tris(propanoyloxy)oxan-2-yl(2E)-3-(1H-indol-3-yl)prop-2-enoate

This compound was prepared following a modified procedure described forcompound 74.

Compound 103:(3S,4S,5R,6S)-6-{[2-(1H-indol-3-yl)acetyl]oxy}-4,5-bis(propanoyloxy)oxan-3-ylpropanoate

This compound was prepared following a modified procedure described forcompound 74.

Compound 104: (2S,3R,4R,5R)-3,4,5-tris(propanoyloxy)oxan-2-yl propanoate

This compound was prepared according to a modified procedure describedfor the preparation of compound 219.

Compound 105: (3R,4R,5R)-2-hydroxy-4,5-bis(propanoyloxy)oxan-3-ylpropanoate

This compound was prepared according to a modified procedure describedfor the preparation of compound 21.

Compound 106:(2S,3R,4R,5R)-2-{[2-(1H-indol-3-yl)acetyl]oxy}-4,5-bis(propanoyloxy)oxan-3-ylpropanoate

This compound was prepared following a modified procedure described forcompound 74.

Compound 107:(2R,3R,4R,5S,6S)-6-methyl-3,4,5-tris(propanoyloxy)oxan-2-yl(2E)-3-(1H-indol-3-yl)prop-2-enoate

This compound was prepared following a modified procedure described forcompound 74.

Compound 108:(3S,4S,5R,6R)-6-{[2-(1H-indol-3-yl)acetyl]oxy}-4,5-bis(propanoyloxy)oxan-3-ylpropanoate

This compound was prepared following a modified procedure described forcompound 74.

Compound 109:[(2R,3R,4S,5R,6R)-6-{[3-(1H-indol-3-yl)propanoyl]oxy}-3,4,5-tris(propanoyloxy)oxan-2-yl]methylpropanoate

This compound was prepared following a modified procedure described forcompound 74.

Compound 110:[(2R,3R,4S,5R)-6-hydroxy-3,4,5-tris(propanoyloxy)oxan-2-yl]methylpropanoate

This compound was prepared according to a modified procedure describedfor the preparation of compound 21.

Compound 111:[(2R,3R,4S,5R,6S)-6-{[2-(1H-indol-3-yl)acetyl]oxy}-3,4,5-tris(propanoyloxy)oxan-2-yl]methylpropanoate

This compound was prepared following a modified procedure described forcompound 74.

Compound 112:[(2R,3R,4S,5R,6S)-6-{[3-(1H-indol-3-yl)propanoyl]oxy}-3,4,5-tris(propanoyloxy)oxan-2-yl]methylpropanoate

This compound was prepared following a modified procedure described forcompound 74. LCMS (M+Na⁺): 598.2. ¹H NMR (400 MHz, DMSO-d₆) δ 10.78 (d,J=2.4 Hz, 1H), 7.50-7.44 (m, 1H), 7.34-7.27 (m, 1H), 7.09-7.00 (m, 2H),6.95 (ddd, J=7.9, 6.9, 1.1 Hz, 1H), 5.99 (d, J=8.3 Hz, 1H), 5.45 (t,J=9.6 Hz, 1H), 5.02-4.89 (m, 2H), 4.27-4.15 (m, 2H), 4.03-3.94 (m, 1H),2.97-2.84 (m, 2H), 2.79-2.63 (m, 2H), 2.34-1.98 (m, 8H), 1.05-0.82 (m,12H).

Compound 113:(3R,4R,5S,6S)-2-hydroxy-6-methyl-4,5-bis(propanoyloxy)oxan-3-ylpropanoate

This compound was prepared according to a modified procedure describedfor the preparation of compound 21.

Compound 114:[(2R,3R,4S,5R,6R)-6-{[2-(1H-indol-3-yl)acetyl]oxy}-3,4,5-tris(propanoyloxy)oxan-2-yl]methylpropanoate

This compound was prepared following a modified procedure described forcompound 74.

Compound 115:(2S,3R,4R,5S,6S)-2-{[2-(1H-indol-3-yl)acetyl]oxy}-6-methyl-4,5-bis(propanoyloxy)oxan-3-ylpropanoate

This compound was prepared following a modified procedure described forcompound 74.

Compound 116:(2S,3S,4R,5R,6S)-6-methyl-3,4,5-tris(propanoyloxy)oxan-2-yl(2E)-3-(1H-indol-3-yl)prop-2-enoate

This compound was prepared following a modified procedure described forcompound 74.

Compound 117:(2R,3R,4S,5R)-2-{[2-(1H-indol-3-yl)acetyl]oxy}-4,5-bis(propanoyloxy)oxan-3-ylpropanoate

This compound was prepared following a modified procedure described forcompound 74. LCMS (M+Na⁺): 498.2. ¹H NMR (400 MHz, Chloroform-d) δ 8.10(s, 1H), 7.63-7.56 (m, 1H), 7.33-7.26 (m, 1H), 7.18 (d, J=2.4 Hz, 1H),7.17-7.06 (m, 2H), 6.21 (d, J=3.6 Hz, 1H), 5.42 (t, J=9.9 Hz, 1H),4.98-4.87 (m, 2H), 3.82 (s, 2H), 3.74 (dd, J=11.1, 5.9 Hz, 1H), 3.40 (t,J=11.0 Hz, 1H), 2.22 (qd, J=7.6, 1.9 Hz, 4H), 1.96 (qd, J=7.5, 4.2 Hz,2H), 1.08-0.99 (m, 6H), 0.88 (t, J=7.6 Hz, 3H).

Compound 118:(2S,3S,4R,5R,6S)-2-{[2-(1H-indol-3-yl)acetyl]oxy}-6-methyl-4,5-bis(propanoyloxy)oxan-3-ylpropanoate

Compound 119: (2R,3R,4R,5S,6S)-3,4,5-tris(butanoyloxy)-6-methyloxan-2-ylbutanoate

This compound was prepared according to a modified procedure describedfor the preparation of compound 219. ¹H NMR (400 MHz, DMSO-d6) δ 6.05(d, J=1.3 Hz, 1H), 5.38 (dd, J=3.5, 1.2 Hz, 1H), 5.31 (dd, J=10.1, 3.4Hz, 1H), 4.89 (t, J=9.9 Hz, 1H), 3.90-3.81 (m, 1H), 2.41-2.36 (m, 2H),2.33-2.22 (m, 4H), 2.15 (td, J=7.2, 1.2 Hz, 2H), 1.65-1.56 (m, 2H),1.55-1.41 (m, 6H), 1.11 (d, J=6.2 Hz, 3H), 0.94 (t, J=7.4 Hz, 3H),0.88-0.80 (m, 9H)

Compound 120: (2S,3R,4R,5S,6S)-3,4,5-tris(butanoyloxy)-6-methyloxan-2-ylbutanoate

This compound was prepared according to a modified procedure describedfor the preparation of compound 219.

Compound 121:(2R,3R,4R,5S,6S)-3,4,5-tris({[2-(1H-indol-3-yl)acetyl]oxy})-6-methyloxan-2-yl2-(1H-indol-3-yl)acetate

This compound was prepared following a modified procedure described forcompound 63.

Compound 122:(2S,3R,4R,5S,6S)-3,4,5-tris({[2-(1H-indol-3-yl)acetyl]oxy})-6-methyloxan-2-yl2-(1H-indol-3-yl)acetate

This compound was prepared following a modified procedure described forcompound 63.

Compound 123:(2S,3S,4R,5R,6S)-6-methyl-3,4,5-tris(propanoyloxy)oxan-2-yl3-(1H-indol-3-yl)propanoate

This compound was prepared following a modified procedure described forcompound 74. LCMS (M+Na⁺): 526.2. ¹H NMR (400 MHz, Chloroform-d) δ 8.08(s, 1H), 7.65-7.57 (m, 1H), 7.35 (dt, J=8.1, 1.0 Hz, 1H), 7.24-7.09 (m,2H), 7.09-7.04 (m, 1H), 6.33 (d, J=2.5 Hz, 1H), 5.36-5.26 (m, 2H),5.26-5.22 (m, 1H), 4.01-3.91 (m, 1H), 3.20-3.08 (m, 2H), 2.84 (t, J=7.1Hz, 2H), 2.44 (qd, J=7.6, 0.9 Hz, 2H), 2.24 (q, J=7.5 Hz, 2H), 2.09 (qd,J=7.6, 1.8 Hz, 2H), 1.18 (t, J=7.6 Hz, 3H), 1.09 (t, J=7.6 Hz, 3H),1.05-0.96 (m, 6H)

Compound 124:(3S,4R,5R,6S)-2-hydroxy-6-methyl-4,5-bis(propanoyloxy)oxan-3-ylpropanoate

This compound was prepared according to a modified procedure describedfor the preparation of compound 21.

Compound 125: (3S,4R,5R,6S)-6-methyl-3,4,5-tris(propanoyloxy)oxan-2-ylpropanoate

This compound was prepared according to a modified procedure describedfor the preparation of compound 219.

Compound 126: (2R,3R,4S,5S)-3,4,5-tris(propanoyloxy)oxan-2-yl3-(1H-indol-3-yl)propanoate

This compound was prepared following a modified procedure described forcompound 74. LCMS (M+Na⁺): 512.2. ¹H NMR (400 MHz, Chloroform-d) δ 8.00(s, 1H), 7.57-7.48 (m, 1H), 7.32-7.25 (m, 1H), 7.19 (s, 1H), 7.17-6.93(m, 3H), 6.28 (d, J=3.1 Hz, 1H), 5.32-5.21 (m, 3H), 3.74 (dd, J=13.2,1.4 Hz, 1H), 3.62 (dd, J=13.2, 1.9 Hz, 1H), 3.11-3.01 (m, 2H), 2.77 (t,J=7.4 Hz, 2H), 2.35 (q, J=7.5 Hz, 2H), 2.19 (q, J=7.6 Hz, 2H), 2.04 (qd,J=7.6, 1.6 Hz, 2H), 1.15-1.00 (m, 6H), 0.94 (t, J=7.6 Hz, 3H)

Compound 127: (3S,4S,5R,6R)-6-hydroxy-4,5-bis(propanoyloxy)oxan-3-ylpropanoate

This compound was prepared according to a modified procedure describedfor the preparation of compound 21.

Compound 128: (2S,3R,4S,5S)-3,4,5-tris(propanoyloxy)oxan-2-yl propanoate

This compound was prepared according to a modified procedure describedfor the preparation of compound 219.

Compound 129: (2R,3R,4S,5R)-2-hydroxy-4,5-bis(propanoyloxy)oxan-3-ylpropanoate

This compound was prepared according to a modified procedure describedfor the preparation of compound 21.

Compound 130: (3R,4S,5R)-3,4,5-tris(propanoyloxy)oxan-2-yl propanoate

This compound was prepared according to a modified procedure describedfor the preparation of compound 219.

Compound 131: [(3S,4S,5R)-2,3,4,5-tetrakis(butanoyloxy)oxan-2-yl]methylbutanoate

This compound was prepared according to a modified procedure describedfor the preparation of compound 219.

Compound 132: (2S,3R,4S,5R)-3,4,5-tris(propanoyloxy)oxan-2-yl3-(1H-indol-3-yl)propanoate

This compound was prepared following a modified procedure described forcompound 74. LCMS: (m+Na+) 512.2. ¹H NMR (400 MHz, DMSO-d6) δ10.89-10.66 (m, 1H), 7.52-7.44 (m, 1H), 7.30 (m, 1H), 7.10-7.01 (m, 2H),6.95 (m, 1H), 5.84 (d, 1H), 5.30 (t, 1H), 4.96-4.84 (m, 2H), 3.97 (dd,1H), 3.67 (dd, 1H), 2.92 (t, 2H), 2.70 (td, 2H), 2.31-2.10 (m, 6H),1.05-0.85 (m, 9H)

Compound 133:[(2R,3R,4S,5R)-3,4,5-tris(butanoyloxy)-6-hydroxyoxan-2-yl]methylbutanoate

Compound 134:[(2R,3R,4S,5R)-3,4,5,6-tetrakis(butanoyloxy)oxan-2-yl]methyl butanoate

Step 1:

To the solution of 2-hydroxybenzoic acid (6 g, 43.44 mmol, 7.50 mL, 1eq) and CDI (8.45 g, 52.13 mmol, 1.2 eq) in DMF (50 mL) was added DBU(7.94 g, 52.13 mmol, 7.86 mL, 1.2 eq) and t-BuOH (6.47 g, 87.32 mmol,8.35 mL, 2.01 eq). The mixture was stirred at 15° C. for 16 h. LCMS(ET14826-364-P1A) showed the reaction was completed. The solvent wasremoved under reduced pressure. The crude product was purified by silicagel chromatography eluted with Petroleum ether/Ethyl acetate=1:0-2:1 togive tert-butyl 2-hydroxybenzoate (5 g, 25.74 mmol, 59.26% yield) ascolorless oil showed by ¹H NMR. LCMS: (M−H⁺): 193.1 @ 1.988 min

Step 2:

To the solution of (2R,3S,4R,5R)-2,3,4,5,6-pentahydroxyhexanal (20 g,111.02 mmol, 1 eq) in DCM (500 mL) was added butyryl chloride (94.63 g,888.12 mmol, 92.77 mL, 8 eg) and the mixture was stirred at 15° C. for0.5 h. Then pyridine (70.25 g, 888.12 mmol, 71.68 mL, 8 eg) was added tothe solution dropwise slowly. After the addition, the mixture wasstirred at 15° C. for another 16 h. LCMS (ET14826-367-P1A) showed thereaction was completed. The solvent was removed under reduced pressure.The crude product was purified by silica gel chromatography eluted withPetroleum ether/Ethyl acetate=1:0-5:1 to give(3R,4S,5R,6R)-6-((butyryloxy)methyl) tetrahydro-2H-pyran-2,3,4,5-tetrayltetrabutyrate (58 g, 109.31 mmol, 98.46% yield) as yellow oil showed by¹H NMR. LCMS: (M+18): 548.3 @ 1.640 min

Step 3:

To the solution of(3R,4S,5R,6R)-6-((butyryloxy)methyl)tetrahydro-2H-pyran-2,3,4,5-tetrayltetrabutyrate (10 g, 18.85 mmol, 1 eg) in THF (85 mL) and H₂O (5 mL) wasadded methanamine/THF (2 M, 12.25 mL, 1.3 eg). Then the mixture wasstirred at 15° C. for 16 h. LCMS (ET14826-370-P1A2) showed most of thestarting material was consumed and the desired MS was detected. Thesolvent was removed under reduced pressure. The crude product waspurified by silica gel chromatography eluted with Petroleum ether/Ethylacetate=10:1-1:1 to give(2R,3R,4S,5R)-2-((butyryloxy)methyl)-6-hydroxytetrahydro-2H-pyran-3,4,5-triyltributyrate (10 g, 21.50 mmol, 57.03% yield, 99% purity) as yellow oil.LCMS: (M+18): 478.3 @ 1.478 min; LCMS: (M+Na⁺): 483.1 @ 3.678, 3.742 min

Compound 135:(3R,4R,5R)-6-hydroxy-4,5-bis({[3-(1H-indol-3-yl)propanoyl]oxy})oxan-3-yl3-(1H-indol-3-yl)propanoate

This compound was prepared following a modified procedure described forcompound 37 with the exception that the synthesis was stopped at thestage producing the title compound.

Compound 136:(3S,4R,5R,6S)-4,5-bis(butanoyloxy)-2-hydroxy-6-methyloxan-3-yl butanoate

This compound was prepared according to a modified procedure describedfor the preparation of compound 21.

Compound 137: (3S,4R,5R,6S)-3,4,5-tris(butanoyloxy)-6-methyloxan-2-ylbutanoate

This compound was prepared according to a modified procedure describedfor the preparation of compound 219.

Compound 138:(3R,4R,5R)-6-hydroxy-4,5-bis({[2-(1H-indol-3-yl)acetyl]oxy})oxan-3-yl2-(1H-indol-3-yl)acetate

This compound was prepared following a modified procedure described forcompound 37 with the exception that the synthesis was stopped at thestage producing the title compound.

Compound 139:(3S,4S,5R)-6-hydroxy-4,5-bis({[2-(1H-indol-3-yl)acetyl]oxy})oxan-3-yl2-(1H-indol-3-yl)acetate

This compound was prepared following a modified procedure described forcompound 37 with the exception that the synthesis was stopped at thestage producing the title compound.

Compound 140:(2S,3R,4R,5R)-3,4,5-tris({[2-(1H-indol-3-yl)acetyl]oxy})oxan-2-yl2-(1H-indol-3-yl)acetate

This compound was prepared following a modified procedure described forcompound 74.

Compound 141:(2S,3S,4R,5R)-4,5-bis(butanoyloxy)-6-hydroxy-2-methyloxan-3-yl butanoate

This compound was prepared according to a modified procedure describedfor the preparation of compound 21.

Compound 142: (3R,4R,5S,6S)-3,4,5-tris(butanoyloxy)-6-methyloxan-2-ylbutanoate

This compound was prepared according to a modified procedure describedfor the preparation of compound 219.

Compound 143:(2S,3S,4R,5R)-6-hydroxy-4,5-bis({[2-(1H-indol-3-yl)acetyl]oxy})-2-methyloxan-3-yl2-(1H-indol-3-yl)acetate

This compound was prepared following a modified procedure described forcompound 37 with the exception that the synthesis was stopped at thestage producing the title compound.

Compound 144:(3S,4S,5R)-6-hydroxy-4,5-bis({[3-(1H-indol-3-yl)propanoyl]oxy})oxan-3-yl3-(1H-indol-3-yl)propanoate

This compound was prepared following a modified procedure described forcompound 37 with the exception that the synthesis was stopped at thestage producing the title compound.

Compound 145:(2R,3R,4R,5R)-3,4,5-tris({[2-(1H-indol-3-yl)acetyl]oxy})oxan-2-yl2-(1H-indol-3-yl)acetate

This compound was prepared following a modified procedure described forcompound 63.

Compound 146:(2S,3R,4S,5S)-3,4,5-tris({[2-(1H-indol-3-yl)acetyl]oxy})oxan-2-yl2-(1H-indol-3-yl)acetate

This compound was prepared following a modified procedure described forcompound 63.

Compound 147:(3R,4R,5S,6S)-3,4,5-tris({[3-(1H-indol-3-yl)propanoyl]oxy})-6-methyloxan-2-yl3-(1H-indol-3-yl)propanoate

This compound was prepared following a modified procedure described forcompound 63.

Compound 148:(2R,3R,4S,5S)-3,4,5-tris({[2-(1H-indol-3-yl)acetyl]oxy})oxan-2-yl2-(1H-indol-3-yl)acetate

This compound was prepared following a modified procedure described forcompound 63.

Compound 149:(2R,3R,4S,5S)-3,4,5-tris({[3-(1H-indol-3-yl)propanoyl]oxy})oxan-2-yl3-(1H-indol-3-yl)propanoate

This compound was prepared following a modified procedure described forcompound 63. LCMS (M+H⁺): 835.3. ¹H NMR (400 MHz, Chloroform-d) δ 7.68(d, J=7.1 Hz, 2H), 7.57 (d, J=10.5 Hz, 2H), 7.54-7.48 (m, 2H), 7.48-7.41(m, 2H), 7.20-6.98 (m, 12H), 6.92 (dd, J=13.6, 2.4 Hz, 2H), 6.60 (d,J=2.3 Hz, 1H), 6.49 (d, J=2.3 Hz, 1H), 6.27 (d, J=3.6 Hz, 1H), 5.32 (dd,J=10.7, 3.5 Hz, 1H), 5.25 (s, 1H), 5.24-5.17 (m, 1H), 3.75 (d, J=13.2Hz, 1H), 3.59 (dd, J=13.2, 2.0 Hz, 1H), 3.13-2.94 (m, 4H), 2.90-2.67 (m,8H), 2.31-2.02 (m, 4H)

Compound 150:(2R,3R,4R,5R)-3,4,5-tris({[3-(1H-indol-3-yl)propanoyl]oxy})oxan-2-yl3-(1H-indol-3-yl)propanoate

This compound was prepared following a modified procedure described forcompound 63. LCMS (M+H⁺): 835.3. ¹H NMR (400 MHz, Chloroform-d) δ 7.79(d, J=15.3 Hz, 3H), 7.69 (s, 1H), 7.53-7.43 (m, 4H), 7.27-7.14 (m, 4H),7.14-7.06 (m, 4H), 7.02 (t, J=7.4 Hz, 4H), 6.82 (s, 2H), 6.76 (s, 2H),5.97 (s, 1H), 5.17 (d, J=4.7 Hz, 1H), 5.06 (t, J=6.1 Hz, 1H), 4.04 (s,1H), 4.01-3.93 (m, 1H), 3.81 (dd, J=12.1, 5.7 Hz, 1H), 3.03-2.88 (m,8H), 2.65-2.56 (m, 6H), 2.44 (t, J=7.5 Hz, 2H)

Compound 151: (3S,4S,5R)-4,5-bis(butanoyloxy)-6-hydroxyoxan-3-ylbutanoate

Compound 152: (2R,3R,4S,5S)-3,4,5-tris(butanoyloxy)oxan-2-yl butanoate

Step 1:

To the solution of (3R,4S,5S)-tetrahydro-2H-pyran-2,3,4,5-tetraol (10 g,66.61 mmol, 1 eq), TEA (53.92 g, 532.87 mmol, 74.17 mL, 8 eq) and DMAP(1.63 g, 13.32 mmol, 0.2 eq) in DCM (100 mL) was added butyric anhydride(52.69 g, 333.05 mmol, 54.48 mL, 5 eq) at 0° C. Then the solution wasstirred 0° C. for 1 h and stirred at 15° C. for another 15 h. TLC showedthe reaction was completed. The solvent was removed under reducedpressure. The crude product was purified by silica gel chromatographyeluted with Petroleum ether/Ethyl acetate=1:0 to give compound 152(3R,4S,5S)-tetrahydro-2H-pyran-2,3,4,5-tetrayl tetrabutyrate (28 g,65.04 mmol, 97.65% yield, 100% purity) as yellow oil. LCMS: (M+Na⁺): 453@ 1.592 min. ¹H NMR (400 MHz, Chloroform-d) δ 6.4 (m, 1H), 5.4-5.3 (m,3H), 4.1-3.8 (m, 2H), 3.7-3.3 (m, 2H), 2.4-2.2 (m, 8H), 1.7-1.6 (m, 8H),1.0-0.9 (m, 12H).

Step 2:

To the solution of compound 152(3R,4S,5S)-tetrahydro-2H-pyran-2,3,4,5-tetrayl tetrabutyrate (25 g,58.07 mmol, 1 eq) in THF (200 mL) and H₂O (10 mL) was addedmethanamine/THF (2 M, 37.75 mL, 1.3 eq). Then the mixture was stirred at15° C. for 16 h. LCMS showed the desired MS. The solvent was removedunder reduced pressure. The crude product was purified by silica gelchromatography eluted with Petroleum ether/Ethyl acetate=1:0-2:1 to givecompound 151 (3R,4S,5S)-2-hydroxytetrahydro-2H-pyran-3,4,5-triyltributyrate (14 g, 38.85 mmol, 33.45% yield) as yellow oil. LCMS:(M+H₂O⁺): 378 @2.833, 2.934 min. ¹H NMR (400 MHz, Chloroform-d) δ5.5-5.4 (m, 1H), 5.4-5.3 (m, 1H), 5.3-5.1 (m, 2H), 4.6 (m, 1H) 4.3-4.0(m, 1H), 3.7-3.6 (m, 1H), 2.4-2.3 (m, 6H), 1.7-1.6 (m, 6H), 1.0-0.9 (m,9H)

Compound 153: (3R,4R,5R)-4,5-bis(butanoyloxy)-2-hydroxyoxan-3-ylbutanoate

This compound was prepared according to a modified procedure describedfor the preparation of compound 21.

Compound 154:[(2R,3R,4S,5R)-4-(butanoyloxy)-5-[(butanoyloxy)methyl]-5-hydroxy-3-{[(2S,3R,4S,5S,6R)-3,4,5-tris(butanoyloxy)-6-[(butanoyloxy)methyl]oxan-2-yl]oxy}oxolan-2-yl]methylbutanoate

This compound was prepared according to a modified procedure describedfor the preparation of compound 219.

Compound 155:[(2R,3R,4S,5S)-4,5-bis(butanoyloxy)-5-[(butanoyloxy)methyl]-3-{[(2S,3R,4S,5S,6R)-3,4,5-tris(butanoyloxy)-6-[(butanoyloxy)methyl]oxan-2-yl]oxy}oxolan-2-yl]methylbutanoate

This compound was prepared according to a modified procedure describedfor the preparation of compound 219.

Compound 156:(3R,4S,5R)-6-hydroxy-4,5-bis[(4-phenylbutanoyl)oxy]oxan-3-yl4-phenylbutanoate

This compound was prepared according to the description in WO2018/226732. ¹H NMR (CDCl₃): δ 7.0-7.2 (m, 15H) 5.5 (dd, 1H), 5.4 (m,1H), 4.8-5.0 (m, 2H), 4.1 (brs, 1H), 3.8, (dd, 2H), 2.5-2.6 (m, 6H),2.2-2.3 (m, 6H), 1.8-0.9 (m, 6H) ppm

Compound 157: (3R,4S,5R)-4,5,6-tris[(4-phenylbutanoyl)oxy]oxan-3-yl4-phenylbutanoate

This compound was prepared according to a modified procedure describedfor the preparation of compound 219.

Compound 158: (3R,4S,5R)-4,5-bis(butanoyloxy)-2-hydroxyoxan-3-ylbutanoate

Step 1

To a solution of pyridine (316.13 g, 4.00 mol, 322.58 mL, 6 eq) in CHCl₃(1 L) was added butanoyl chloride (425.83 g, 4.00 mol, 417.48 mL, 6 eq)and DMAP (2.44 g, 19.98 mmol, 0.03 eq) at 0° C.(2R,3S,4R)-2,3,4,5-tetrahydroxypentanal (100 g, 666.09 mmol, 1 eq) wasadded into the mixture at 0° C. and the mixture was stirred at 25° C.for 12 h. TLC showed the starting reactant consumed. The mixturereaction was concentrated. The residue was purified by columnchromatography (SiO₂, Petroleum ether/Ethyl acetate=20/1 to 10:1 to3/1). [(3R,4S,5R)-4,5,6-tri(butanoyloxy)tetrahydropyran-3-yl]butanoate(165 g, 325.79 mmol, 48.91% yield, 85% purity) was obtained as colorlessoil.

Step 2

To a solution of [(3R,4S,5R)-4,5,6-tri(butanoyloxy)tetrahydropyran-3-yl]butanoate (55 g, 127.76 mmol, 1 eq) in THF (800 mL) was added MeNH₂ inH₂O (14.88 g, 191.64 mmol, 40% purity, 1.5 eq). The mixture was stirredat 25° C. for 12 h. TLC showed the starting reactant consumed. Themixture reaction was concentrated. The residue was purified by columnchromatography (SiO₂, Petroleum ether/Ethyl acetate=10/1 to 5:1).[(3R,4S,5R)-4,5-di(butanoyloxy)-6-hydroxy-tetrahydropyran-3-yl]butanoate (50 g, 124.86 mmol, 48.87% yield, 90% purity) was obtained.The compound was combined with other batches. In total, 99 g wasobtained as a yellow solid. LCMS: (M+Na⁺): 383.1 @ 3.490 min. ¹H NMR(400 MHz, DMSO-de) δ 7.2-7.0 (m, 1H), 5.4-5.1 (m, 2H), 4.9-4.7 (m, 2H),3.7-3.3 (m, 2H), 2.4-2.2 (m, 6H), 1.5-1.4 (m, 6H), 0.9-0.8 (m, 9H)

Compound 159: (2R,3R,4S,5R)-3,4,5-tris(butanoyloxy)oxan-2-yl butanoate

This compound was prepared according to a modified procedure describedfor the preparation of compound 219.

Compound 160: (3R,4R,5R)-3,4,5-tris(butanoyloxy)oxan-2-yl butanoate

This compound was prepared according to a modified procedure describedfor the preparation of compound 219.

Compound 161:(2R,3R,4S,5S,6S)-2-(acetoxymethyl)-6-methoxytetrahydro-2H-pyran-3,4,5-triyltriacetate

This compound was prepared according to a modified procedure describedfor the preparation of compound 219.

LCMS: (m+Na+) 385.1. ¹H NMR (400 MHz, DMSO-d6) δ 5.16-5.07 (m, 3H), 4.79(d, 1H), 4.17 (dd, 1H), 4.07 (dd, 1H), 3.95-3.87 (m, 1H), 3.36 (s, 3H),2.12 (s, 3H), 2.04 (d, 6H), 1.95 (s, 3H)

Compound 162:(2S,3R,4R,5S,6R)-6-(acetoxymethyl)-3-aminotetrahydro-2H-pyran-2,4,5-triyltriacetate

Stir Boc-protected glucosamine in acetic anhydride and purify. Then,deprotect with HCl in dioxane to yield the title compound as the HClsalt. LCMS: (M+Na+): 370.1. ¹H NMR (400 MHz, DMSO-d₆) δ 8.94-8.54 (m,3H), 6.01-5.82 (m, 1H), 5.42-5.27 (m, 1H), 4.94 (t, J=9.6 Hz, 1H), 4.20(dd, J=12.5, 4.4 Hz, 1H), 4.13-3.92 (m, 2H), 3.67-3.51 (m, 1H), 2.18 (s,3H), 2.08-1.95 (m, 9H)

Compound 163:(2S,3S,4S,5R,6R)-3,4,5,6-tetraacetoxytetrahydro-2H-pyran-2-carboxylicacid

This compound was prepared following a modified procedure described forcompounds 46 and 47. LCMS: (M+NH₄ ⁺): 380.1. ¹H NMR (400 MHz,Chloroform-d) δ 6.40 (s, 1H), 5.52 (t, J=9.5 Hz, 1H), 5.32-5.22 (m, 1H),5.16-5.08 (m, 1H), 4.46 (d, J=9.7 Hz, 1H), 2.20 (s, 3H), 2.08-2.00 (m,9H).

Compound 164:(2S,3S,4S,5R,6S)-3,4,5,6-tetraacetoxytetrahydro-2H-pyran-2-carboxylicacid

This compound was prepared following a modified procedure described forcompounds 46 and 47. LCMS: (M+NH4+): 380.0. ¹H NMR (400 MHz,Chloroform-d) δ 5.81 (d, J=7.6 Hz, 1H), 5.39-5.26 (m, 2H), 5.22-5.10 (m,1H), 4.32-4.22 (m, 1H), 2.14 (s, 3H), 2.12-2.03 (m, 9H).

Compound 165:2-[3,4-bis(propanoyloxy)phenyl]-3-hydroxy-4-oxo-7-(propanoyloxy)-4H-chromen-5-ylpropanoate

This compound was prepared following a modified procedure described forcompound 26. ¹H NMR (400 MHz, Chloroform-d) δ 12.10 (s, 1H), 7.88-7.58(m, 2H), 7.36 (d, J=8.3 Hz, 1H), 6.86 (d, J=2.0 Hz, 1H), 6.59 (d, J=2.0Hz, 1H), 2.87-2.37 (m, 8H), 1.41-1.12 (m, 12H)

Compound 166: 4-[3,5,7-tris(butanoyloxy)-4-oxo-4H-chromen-2-yl]phenylbutanoate

To a mixture of 3,5,7-trihydroxy-2-(4-hydroxyphenyl)chromen-4-one (500mg, 1.75 mmol, 1 eg), TEA (883.80 mg, 8.73 mmol, 1.22 mL, 5 eg) in THF(20 mL) was added butanoyl chloride (930.62 mg, 8.73 mmol, 912.37 uL, 5eg) slowly at 0° C. And then the mixture was stirred at 50° C. for 5 hrunder N2 atmosphere. LC-MS showed reactant was consumed completely andone main peak with desired mass was detected. The reaction mixture wasquenched by addition H₂O 200 mL at 25° C. and then extracted with EtOAc180 mL (60 mL*3). The combined organic layers were washed with brine 20mL, dried over Na₂SO₄, filtered and concentrated under reduced pressureto give a residue. The residue was purified by column chromatography(SiO₂, Petroleum ether/Ethyl acetate=5/1 to 3:1).[4-[3,5,7-tri(butane-yloxy)-4-oxo-chromen-2-yl]phenyl]butanoate (437 mg,734.02 umol, 42.02% yield, 95.17% purity) was obtained as a white solid.LCMS: (M+H⁺) 567.2 @ 1.577 min; LCMS: (M+H⁺) 567.2 @ 3.520 min. ¹H NMR(400 MHz, Chloroform-d) δ 12.10 (s, 1H), 7.88-7.58 (m, 2H), 7.36 (d,J=8.3 Hz, 1H), 6.86 (d, J=2.0 Hz, 1H), 6.59 (d, J=2.0 Hz, 1H), 2.87-2.37(m, 8H), 1.41-1.12 (m, 12H)

Compound 167: 4-[4-oxo-3,5,7-tris(propanoyloxy)-4H-chromen-2-yl]phenylpropanoate

This compound was prepared following a modified procedure described forcompound 27. LCMS: (M+H⁺) 511.2. ¹H NMR (400 MHz, Chloroform-d) δ7.88-7.80 (m, 2H), 7.32 (d, J=2.2 Hz, 1H), 7.28-7.21 (m, 2H), 6.86 (d,J=2.2 Hz, 1H), 2.77 (q, J=7.5 Hz, 2H), 2.68-2.58 (m, 6H), 1.33-1.23 (m,9H), 1.21 (t, J=7.5 Hz, 3H)

Compound 168: 5-hydroxy-4-oxo-2-[4-(propanoyloxy)phenyl]-4H-chromen-7-ylpropanoate

Propionic anhydride (641 uL, 5.03 mmol) was added dropwise to a stirredsolution of 5,7-dihydroxy-2-(4-hydroxyphenyl)-4H-chromen-4-one (170 mg,0.63 mmol) in 1 mL pyridine at 0° C. under nitrogen. The reaction wasstirred at room temperature for 16 hours then diluted with 20 mL ethylacetate. The organic layer was washed with 10 mL 1M HCl twice followedby brine, dried over MgSO4. filtered and concentrated. The residue wasdissolved in DMSO and purified by reverse phase flash chromatography(10-90% acetonitrile in water). Fraction was concentrated bylyophilization to yield5-hydroxy-4-oxo-2-[4-(propanoyloxy)phenyl]-4H-chromen-7-yl propanoate(48 mg, 20% yield) as a white solid, LCMS: (M+H) 383.1. ¹H NMR (400 MHz,DMSO-d6) δ 12.83 (s, 1H), 8.21-8.15 (m, 2H), 7.40-7.34 (m, 2H), 7.14 (s,1H), 7.11 (d, 1H), 6.68 (d, 1H), 2.65 (qd, 4H), 1.16 (t, 6H)

Compound 169:3,5-bis(butanoyloxy)-4-oxo-2-[3,4,5-tris(butanoyloxy)phenyl]-4H-chromen-7-ylbutanoate

A mixture of 3,5,7-trihydroxy-2-(3,4,5-trihydroxyphenyl)chromen-4-one(0.2 g, 628.47 umol, 1 eq), butanoyl butanoate (795.36 mg, 5.03 mmol,822.50 uL, 8 eq) in Pyridine (5 mL) was degassed and purged with N₂ for3 times, and then the mixture was stirred at 20° C. for 12 hr under N₂atmosphere. TLC indicated the reaction was completed and one new spotformed. The reaction mixture was washed with H₂O (5 mL), filtered andthe filter cake was concentrated under reduced pressure to give aresidue. Compound[3,5-di(butanoyloxy)-4-oxo-2-[3,4,5-tri(butanoyloxy)phenyl]chromen-7-yl]butanoate (0.378 g, 501.43 umol, 79.79% yield, 98% purity) was obtainedas off-white solid. LCMS: (M+H⁺) 739.2 @3.587 min. ¹H NMR (400 MHz,Chloroform-d) δ 7.62 (s, 2H), 7.35 (d, J=2.2 Hz, 1H), 6.88 (d, J=2.2 Hz,1H), 2.74 (t, J=7.5 Hz, 2H), 2.69-2.52 (m, 10H), 1.91-1.70 (m, 12H),1.12-0.98 (m, 18H).

Compound 170:5-(butanoyloxy)-2-[4-(butanoyloxy)phenyl]-4-oxo-4H-chromen-7-ylbutanoate

To a solution of 5,7-dihydroxy-2-(4-hydroxyphenyl)chromen-4-one (500 mg,1.85 mmol, 1 eq) in Py. (5 mL) was added butanoyl butanoate (1.76 g,11.10 mmol, 1.82 mL, 6 eq) at 25° C. The mixture was stirred at 25° C.for 12 hr. LCMS showed the desired compound was detected. The reactionmixture was concentrated under reduced pressure to give a residue. Theresidue were washed with H₂O (20 mL) and petroleum ether (20 mL), andconcentrated under reduced pressure to give a residue.

Compound [4-[5,7-di(butanoyloxy)-4-oxo-chromen-2-yl]phenyl] butanoate(167 mg, 340.60 umol, 18.41% yield, 98% purity) was obtained as a yellowsolid. LCMS: (M+H⁺) 481.1 @ 3.244 min. ¹H NMR (400 MHz, Chloroform-d) δ7.92-7.83 (m, 2H), 7.34 (d, J=2.3 Hz, 1H), 7.28-7.22 (m, 2H), 6.83 (d,J=2.2 Hz, 1H), 6.62 (s, 1H), 2.72 (t, J=7.5 Hz, 2H), 2.58 (td, J=7.4,1.8 Hz, 4H), 1.87-1.74 (m, 6H), 1.12-1.02 (m, 9H)

Compound 171:2-[3,4-bis(butanoyloxy)phenyl]-5-(butanoyloxy)-4-oxo-4H-chromen-7-ylbutanoate

To a solution of 2-(3,4-dihydroxyphenyl)-5,7-dihydroxy-chromen-4-one(500 mg, 1.75 mmol, 1 eq) in Pyridine (10 mL) was added butanoylbutanoate (2.21 g, 13.97 mmol, 2.29 mL, 8 eg) at 25° C. The mixture wasstirred at 25° C. for 12 hr. LCMS showed the desired compound wasdetected. The reaction mixture was concentrated under reduced pressureto give a residue. The residue were washed with H₂O (20 mL) andpetroleum ether (20 mL), and concentrated under reduced pressure to givea residue. Compound[2-butanoyloxy-4-[5,7-di(butanoyloxy)-4-oxo-chromen-2-yl]phenyl]butanoate (155 mg, 270.83 umol, 15.50% yield, 99% purity) was obtainedas a yellow solid. LCMS: (M+H⁺) 567.1 @3.385 min. ¹H NMR (400 MHz,Chloroform-d) δ 7.76-7.66 (m, 2H), 7.40-7.32 (m, 2H), 6.83 (d, J=2.2 Hz,1H), 6.60 (s, 1H), 2.72 (t, J=7.5 Hz, 2H), 2.62-2.51 (m, 6H), 1.91-1.72(m, 8H), 1.11-1.01 (m, 12H).

Compound 172:4-oxo-7-(propanoyloxy)-2-[4-(propanoyloxy)phenyl]-4H-chromen-5-ylpropanoate

To a solution of 5,7-dihydroxy-2-(4-hydroxyphenyl)chromen-4-one (500 mg,1.85 mmol, 1 eq) in Pyridine (5 mL) was added propanoyl propanoate (1.44g, 11.10 mmol, 1.43 mL, 6 eq) at 25° C. The mixture was stirred at 25°C. for 12 hr. LCMS showed the desired compound was detected. Thereaction mixture was concentrated under reduced pressure to give aresidue. The residue were washed with H₂O (20 mL) and petroleum ether(20 mL), and concentrated under reduced pressure to give a residue.Compound [4-[4-oxo-5,7-di(propanoyloxy)chromen-2-yl]phenyl] propanoate(156 mg, 351.73 umol, 19.01% yield, 98.85% purity) was obtained as ayellow solid. LCMS: (M+H⁺) 439.1 @ 3.130 min. ¹H NMR (400 MHz,Chloroform-d) δ 7.94-7.86 (m, 2H), 7.38 (d, J=2.2 Hz, 1H), 7.32-7.24 (m,2H), 6.87 (d, J=2.2 Hz, 1H), 6.64 (s, 1H), 2.80 (q, J=7.5 Hz, 2H), 2.66(qd, J=7.5, 1.8 Hz, 4H), 1.38-1.26 (m, 9H)

Compound 173:[4-[4-oxo-5,7-di(propanoyloxy)chromen-2-yl]-2-propanoyloxy-phenyl]propanoate

Propionic anhydride (1.33 mL, 10.4 mmol) was added dropwise to a stirredsolution of luteolin (0.3 g, 1.04 mmol) in anhydrous pyridine (2.5 mL,31.2 mmol) at 0° C. under N2 atmosphere. The resulting stirred solutionwas allowed to come to room temperature and reaction was monitored tocompletion by LCMS. The solution was diluted with 30 mL ethyl acetateand washed with H₂O (30 mL), 1M HCl (30 mL), H₂O (30 mL), and saturatedNaHCO₃ (30 mL). The organic layer was dried over sodium sulfate,filtered, and concentrated by rotary evaporation. The crude residue waspurified by flash chromatography (silica, 10-100% ethyl acetate inhexanes) and fractions were concentrated by rotary evaporation to yieldcompound 173 (0.073 g, 15% yield) as an off-white solid. 1H-NMR(DMSO-d6, 400 MHz): δ 12.75 (s, 1H), 8.07 (m, 2H), 7.5 (m, 1H), 7.15 (s,1H), 7.12 (d, 1H), 6.66 (d, 1H), 2.59-2.66 (m, 6H), 1.11-1.17 (m, 9H)

Compound 174:[4-[4-oxo-3,5,7-tri(propanoyloxy)chromen-2-yl]-2-propanoyloxy-phenyl]propanoate

Propionic anhydride (2.1 mL, 16.5 mmol) was added dropwise to a stirredsolution of quercetin (0.5 g, 1.65 mmol) in anhydrous pyridine (3.98 mL,49.5 mmol) at 0° C. under N2 atmosphere. The resulting stirred solutionwas allowed to come to room temperature and reaction was monitored tocompletion by LCMS. The solution was diluted with 30 mL ethyl acetateand washed with H₂O (30 mL), 1M HCl (30 mL), H₂O (30 mL), and saturatedNaHCO₃ (30 mL). The organic layer was dried over sodium sulfate,filtered, and concentrated by rotary evaporation. The crude residue waspurified by flash chromatography (silica, 10-100% ethyl acetate inhexanes) and fractions were concentrated by rotary evaporation to yieldCompound 174 (0.1 g, 10% yield) as a white solid. ¹H NMR (DMSO-d6, 400MHz): δ 7.85 (m, 2H), 7.66 (d, 1H), 7.54 (d, 1H), 7.18 (d, 1H),2.62-2.89 (m, 10H), 1.09-1.19 (m, 20H)

Compound 175: (S)-2-(4-(butyryloxy)phenyl)-4-oxochromane-5,7-diyldibutyrate

To a solution of 5,7-dihydroxy-2-(4-hydroxyphenyl)chroman-4-one (0.500g) in pyridine (10 mL), was added butanoyl butanoate (1.02 g). Thereaction mixture was stirred at 15° C. for 12 h. The mixture wasconcentrated. The residue was purified by column chromatography (SiO₂,petroleum ether/ethyl acetate gradient) to give compound 175 (0.325 g,34% yield) as a white solid. LCMS: 500.2 (M+H₂O⁺) ¹H NMR (400 MHz,CDCl₃) δ 7.463 (d, 2H), 7.158 (d, 2H), 6.786 (d, 1H), 6.536 (d, 1H),5.483 (m, 1H), 3.031 (m, 1H), 2.662 (m, 1H), 2.586-2.524 (m, 6H),1.837-1.785 (m, 6H), 1.089-1.021 (m, 9H)

Compound 176: (S)-2-(4-acetoxyphenyl)-4-oxochromane-5,7-diyl diacetate

5,7-dihydroxy-2-(4-hydroxyphenyl)chroman-4-one (0.500 g) was dissolvedwith pyridine (10 mL), and then acetyl acetate (0.844 g) was added tothe mixture reaction. The reaction mixture was stirred at 15° C. for 12h. The mixture reaction was concentrated under reduced pressure. Theresidue was purified by column chromatography (SiO₂, petroleumether/ethyl acetate gradient) to give compound 176 (0.300 g, 39% yield)as a white solid. LCMS: 416.1 (M+H₂O⁺) ¹H NMR (400 MHz, CDCl₃) δ 7.468(d, 2H), 7.166 (d, 2H), 6.793 (d, 1H), 6.551 (d, 1H), 5.497 (dd, 1H),3.039 (dd, 1H), 2.783 (dd, 1H), 2.393 (s, 3H), 2.326 (S, 3H), 2.308 (s,3H).

Compound 177: [4-(3,5,7-triacetoxy-4-oxo-chromen-2-yl)phenyl] acetate

To a mixture of 3,5,7-trihydroxy-2-(4-hydroxyphenyl)chromen-4-one (2 g)in pyridine (15 mL) was added acetyl acetate (30 g), and then themixture was stirred at 15° C. for 12 hr under N₂ atmosphere. The solventwas removed under reduced pressure and the residue was poured intocrushed ice with vigorous stirring. The solid precipitate was collectedby filtration and washed with cold water and then with methanol.Compound 177 (2.1 g, 65% yield) was obtained as a white solid. LCMS:455.0 (M+H⁺) ¹H NMR (400 MHz, CDCl₃) δ 7.858 (d, 2H), 7.339 (d, 1H),7.278-7.257 (m, 2H), 6.883 (d, 1H), 2.447 (s, 3H), 2.357 (S, 6H), 2.333(s, 3H)

Compound 178:[3,5-diacetoxy-4-oxo-2-(3,4,5-triacetoxyphenyl)chromen-7-yl] acetate

To a solution of3,5,7-trihydroxy-2-(3,4,5-trihydroxyphenyl)chromen-4-one (1 g) inpyridine (10 mL) was added acetyl acetate (15.26 g), then the mixturewas stirred at 15° C. for 16 h. The solvent was removed and the mixturewas poured into ice water under stirring. The solid was filtered, washedwith water and dried in vacuum to give compound 178 (1.1 g, 61% yield)as a gray solid. LCMS 571.1 (M+H⁺) 15 ¹H NMR (400 MHz, CDCl₃) δ 7.260(s, 2H), 7.349 (d, 1H), 6.886 (d, 1H), 2.441 (s, 3H), 2.372 (s, 3H),2.353 (s, 3H), 2.341 (s, 3H), 2.333 (s, 6H)

Compound 179:[2-decanoyloxy-4-[3,5,7-tris(decanoyloxy)-4-oxo-chromen-2-yl] phenyl]decanoate

To a mixture of 2-(3,4-dihydroxyphenyl)-3,5,7-trihydroxy-chromen-4-one(1 g) and decanoyl chloride (6.31 g) in THF (50 mL) was added TEA (3.35g) at 25° C., then the mixture was stirred at 55° C. for 12 h. A portionof the solvent was removed in vacuum and the precipitate was collectedby filtration to give compound 179 (2.47 g, 69%) as a white solid. ¹HNMR (400 MHz, CDCl₃). δ 7.772-7.669 (m, 2H), 7.343-7.321 (m, 2H), 6.685(s, 1H), 2.736 (t, 2H), 2.610-2.551 (m, 8H), 1.762 (m, 10H), 1.557-1.295(m, 50H), 0.899 (m, 15H)

Compound 180:[2-octanoyloxy-4-[3,5,7-tri(octanoyloxy)-4-oxo-chromen-2-yl] phenyl]octanoate

To a mixture of 2-(3,4-dihydroxyphenyl)-3,5,7-trihydroxy-chromen-4-one(0.32 g) and octanoyl chloride (1.72 g) in THF (20 mL) was added TEA(1.07 g) at 25° C. Then the mixture was stirred at 55° C. for 12 h. Aportion of the solvent was removed in vacuum and the precipitate wascollected by filtration to give compound 180 (0.20 g, 20%) as a whitesolid. ¹H NMR (400 MHz, CDCl₃). δ 7.709-7.655 (m, 2H), 7.329-7.301 (m,2H), 6.837 (s, 1H), 2.723 (t, 2H), 2.612-2.539 (m, 8H), 1.751 (m, 10H),1.412-1.309 (m, 40H), 0.896 (m, 15H).

Compound 181:[2-butanoyloxy-4-[3,5,7-tri(butanoyloxy)-4-oxo-chromen-2-yl]phenyl]butanoate

To a mixture of 2-(3,4-dihydroxyphenyl)-3,5,7-trihydroxy-chromen-4-one(1 g) and butanoyl chloride (3.53 g) in THF (40 mL) was addedtriethylamine (TEA) (3.35 g) at 25° C., then the mixture was stirred at55° C. for 12 h. The reaction mixture was concentrated in vacuum andpurified by reverse phase prep-HPLC (C18, water (0.05% HCl)-ACNgradient) to give compound 181 (1.13 g, 52% yield) as a colorless solid.LCMS: 653.3 (M+H⁺) ¹H NMR (400 MHz, CDCl₃). δ 7.666-7.608 (m, 2H),7.292-7.210 (m, 2H), 6.880 (s, 1H), 2.542 (t, 2H), 2.535-2.484 (m, 8H),1.753 (m, 10H), 1.020-0.997 (m, 12H), 0.949 (t, 3H).

Compound 182: [2-acetoxy-4-(3,5,7-triacetoxy-4-oxo-chromen-2-yl)phenyl]acetate

To a mixture of 2-(3,4-dihydroxyphenyl)-3,5,7-trihydroxy-chromen-4-one(1 g) and acetic anhydride (2.36 g) in THF (40 mL) was added K₂CO₃ (3.2g) at 25° C., then the mixture was stirred at 55 5° C. for 12 h.Additional acetic anhydride was added (3 equiv.) and the mixture andstirred for another 3 h. The reaction mixture was concentrated in vacuumand purified by reverse phase prep-HPLC (C18; water (0.05% HCl)-ACNgradient) to give compound 182 (0.837 g, 49%) as a white solid. LCMS:513.2 (M+H⁺) ¹H NMR (400 MHz, CDCl₃). δ 7.742-7.703 (m, 2H), 7.373-7.346(m, 2H), 6.888 (s, 1H), 2.443, (s, 3H), 2.356 (s, 6H), 2.350 (s, 6H).

Compound 183:5-amino-2-[(2S,3R,4S,5R)-3,4,5-tri(butanoyloxy)tetrahydropyran-2-yl]oxy-benzoicacid

Step 1

5-Amino salicylic acid (10.0 g) was dissolved in a mixture of dioxane(100 mL), water (100 mL), and NaOH (2.60 g), and the resulting solutionwas cooled in an ice-bath. Di-tert-butyl dicarbonate (Boc anhydride)(15.60 g) was added, and the mixture was warmed to room temperature andstirred for 1.0 h. The solution was concentrated to 60 mL, diluted withethyl acetate (100 mL), and the resulting mixture was cooled in anice-bath. The mixture was acidified with aq. KHSO₄ to pH 2-3. Theaqueous layer was extracted with EtOAc. The organic phase was washedwith water, brine, dried over Na₂SO₄, filtered, and concentrated toafford 5-(tert-butoxycarbonylamino)-2-hydroxy-benzoic acid (7.0 g, 42%).

Step 2

5-(tert-butoxycarbonylamino)-2-hydroxy-benzoic acid (3 g) was dissolvedin DMF, and the resulting solution was cooled to 0° C.1,1′-Carbonyldiimidazole (CDI) was added, and the mixture was stirred atroom temperature for 2 h. Then, tert-butylalcohol (1.7 g) and DBU (2.1g) were added. The reaction was stirred at room temperature overnight.The reaction mixture was poured onto ice-water, and the solid product,tert-butyl 5-(tert-butoxycarbonylamino)-2-hydroxy-benzoate, wascollected by filtration (3.0 g, 81.9%).

Step 3

To a mixture of tert-butyl5-(tert-butoxycarbonylamino)-2-hydroxy-benzoate,[(3R,4S,5R)-4,5-di(butanoyloxy)-6-hydroxy-tetrahydropyran-3-yl]butanoate (1.2 g) and triphenylphosphene (1.2 g) in THF (50 mL) wasadded di-t-butyl azodicarboxylate (DTAD) (1.1 g), and the mixture wasstirred overnight at room temperature. The product was purified byreverse phase chromatography using acetonitrile-water to affordtert-butyl5-(tert-butoxycarbonylamino)-2-[(3R,4S,5R)-3,4,5-tri(butanoyloxy)tetrahydropyran-2-yl]oxy-benzoateas sticky solid (0.6 g, 30%).

Step 4

Tert-butyl5-(tert-butoxycarbonylamino)-2-[(3R,4S,5R)-3,4,5-tri(butanoyloxy)tetrahydropyran-2-yl]oxy-benzoate(600 mg) was added to 4M HCl in dioxane (15 mL) and stirred at roomtemperature overnight. After the consumption of the starting material,the organic phase was evaporated, and the residue was co-evaporated withheptane and dichloromethane twice more. The solid obtained was driedunder high vacuum to afford compound the title product as dark brownsolid (200 mg, 43.8%). Fractionation of the product afforded twoanomeric isomers (compounds 185 and 186). ¹H NMR (DMSO d6): Isomer 1: δ7.62 (d, 1H), 7.45 (dd, 1H), 7.38 (d, 1H), 6 (d, 1H), 5.6 (t, 1H),5.0-5.1 (m, 1H), 4.7-4.75 (m, 1H), 3.6-3.8 (m, 1H), 3.45-3.6 (1H),2.1-2.3 (m, 6H), 1.4-1.6 (m, 6H), 0.75-0.85 (m, 9H). Isomer 2: δ 7.82(d, 1H), 7.5 (dd, 1H), 7.05 (d, 1H), 5.5 (d, 1H), 5.3 (t, 1H), 5.1-5.15(m, 1H), 4.9-5.0 (m, 1H), 4.0-4.08 (m, 1H), 3.7-3.8 (1H), 2.1-2.3 (m,6H), 1.4-1.6 (m, 6H), 0.75-0.85 (m, 9H) ppm

Compound 184:5-amino-2-[(2R,3R,4S,5R)-3,4,5-tri(butanoyloxy)tetrahydropyran-2-yl]oxy-benzoicacid

This compound was prepared according to a modified procedure describedfor the preparation of compound 183. LCMS [M−H]⁻: 494.5. ¹H NMR (400MHz, DMSO-d₆) δ 6.83-6.75 (m, 2H), 6.64-6.53 (m, 1H), 5.58 (d, J=3.6 Hz,1H), 5.53 (t, J=9.9 Hz, 1H), 5.03-4.90 (m, 2H), 3.89 (t, J=10.9 Hz, 1H),3.73 (dd, J=10.9, 5.9 Hz, 1H), 2.38-2.12 (m, 6H), 1.58-1.39 (m, 6H),0.92-0.76 (m, 9H).

Compound 185:5-amino-2-(((2R,3R,4R,5R)-3,4,5-tris(butyryloxy)tetrahydro-2H-pyran-2-yl)oxy)benzoicacid Step 1. Ribose Tetrabutyrate

To a stirred solution of D-(+)-ribose 1 (5 g) in anhydrous pyridine(24.2 mL) was added solution of butyryl chloride (23.70 g) indichloromethane (50 mL) at 0-5° C. The reaction mixture was brought toroom temperature and stirred for 16 h. The mixture was diluted withdichloromethane (100 ml) and washed successively with water (100 mL), 2Naqueous HCl (300 mL), saturated sodium bicarbonate solution (300 mL) andbrine (100 mL). The organic layer was dried over sodium sulfate andconcentrated under reduced pressure. The residue was purified by columnchromatography over silica gel (5-10% EtOAc-hexane gradient) to affordribose tetrabutyrate as a colorless oil (7.5 g, 52%, mixture of α/βanomers).

Step 2. Ribose Tributyrate

Ammonium hydroxide (11 mL) was added slowly to a mixture of ribosetetrabutyrate 2 (7.5 g) in acetonitrile (60 mL) at room temperature andthe resulting reaction mixture was stirred for 5 h. The mixture wasdiluted with MTBE (75 mL) and stirred for 15 minutes. The organic layerwas separated and concentrated under reduced pressure and the residuewas partitioned between MTBE (100 mL) and water (75 mL). The MTBE layerwas separated, dried over sodium sulfate and concentrated under reducedpressure. The residue was purified by column chromatography [usingsilica gel 100-200 mesh and 10-20% EtOAc-Hexane as eluting solvent] toafford ribose tributyrate as a colorless oil (1.1 g, 17%).

Step 3. 5-tert-butoxycarbonylamino-2-hydroxy-benzoic acid

To the stirred solution of 5-amino salicylic acid 4 (5 g) in 1,4-dioxaneand water (1:1; 100 mL) was added NaOH (1.3 g) and Boc-anhydride (7.83g) at 0° C. and the resulting reaction mixture was stirred at roomtemperature for 1 h. The reaction mixture was concentrated under reducedpressure, the residue was diluted with EtOAc (50 mL) and the pH wasadjusted to 3-4 by dropwise addition of 0.5N aqueous HCl at 0° C. Theorganic layer was separated, and the aqueous layer was extracted withEtOAc (50 mL). The combined organic layer was dried over sodium sulfateand concentrated under reduced pressure to provide5-tert-butoxycarbonylamino-2-hydroxy-benzoic acid as off white solid(5.3 g, 64%).

Step 4. 5-tert-butoxycarbonylmethyl-2-hydroxy-benzoic acid tert-butylester

To a stirred solution of 5-tert-butoxycarbonylamino-2-hydroxy-benzoicacid 5 (5.3 g) in DMF (50 mL) was added CDI (3.39 g) at 0-5° C. and themixture was stirred for 2 h. tert-Butanol (4.025 mL) and DBU (2.54 mL)were then added and the mixture was stirred at room temperature for 16h. The mixture was diluted with water (100 mL) and extracted with EtOAc(200 mL). The organic layer was separated, dried over sodium sulfate andconcentrated under reduced pressure. The residue was purified by columnchromatography using silica gel [100-200 mesh; under gradient elution of5-10% EtOAc-Hexane] to afford5-tert-butoxycarbonylmethyl-2-hydroxy-benzoic acid tert-butyl ester asoff white solid (2 g, 31%).

Step 5.(2R,3R,4R,5R)-2-(2-(tert-butoxycarbonyl)-4-((tert-butoxycarbonyl)amino)phenoxy)tetrahydro-2H-pyran-3,4,5-triyltributyrate

hydroxy-benzoic acid tert-butyl ester 6 (0.850 g) and ribose tributyrate(1.04 g) in THF (5 mL) was sequentially added triphenylphosphine (1.03g) and di-tert-butyl azodicarboxylate (0.948 g) at room temperature andthe mixture was stirred for 16 h. The mixture was concentrated underreduced pressure and the residue was purified by column chromatographyover silica gel (5 to 18% EtOAc-Hexane gradient) to afford of crude(2R,3R,4R,5R)-2-(2-(tert-butoxycarbonyl)-4-((tert-butoxycarbonyl)amino)phenoxy)tetrahydro-2H-pyran-3,4,5-triyltributyrate (1.3 g) which was used directly in the next step.

Step 6.5-amino-2-[(2R,3R,4R,5R)-3,4,5-tri(butanoyloxy)tetrahydropyran-2-yl]oxy-benzoicacid

To a stirred solution of crude(2R,3R,4R,5R)-2-(2-(tert-butoxycarbonyl)-4-((tert-butoxycarbonyl)amino)phenoxy)tetrahydro-2H-pyran-3,4,5-triyltributyrate (1.3 g, crude from above experiment) in 1,4-dioxane (7 mL)was added 4N HCl in 1,4-dioxane (10 mL) at 0° C. and the resultingreaction mixture was stirred at room temperature for 16 h. Then reactionmixture was concentrated under reduced pressure and the residue waspurified by reverse phase prep-HPLC to provide5-amino-2-[(2R,3R,4R,5R)-3,4,5-tri(butanoyloxy)tetrahydropyran-2-yl]oxy-benzoicacid (0.05 g). LCMS: 496.5 (M+H⁺) ¹H NMR (400 MHz, DMSO-d6): δ6.919-6.898 (m, 2H), 6.658 (m, 1H), 5.431 (m, 1H), 5.350 (m, 1H), 5.234(m, 1H), 5.161 (m, 1H), 4.213 (m 1H), 3.749 (m, 1H), 2.497-2.268 (m,4H), 2.197 (m, 1H), 1.620-1.487 (m, 6H), 0.926-0.888 (m, 9H) ppm

Compound 186:2-{[(2S,3R,4S,5R)-3,4,5-tris(butanoyloxy)oxan-2-yl]oxy}benzoic acid

Compound 187:2-{[(2R,3R,4S,5R)-3,4,5-tris(butanoyloxy)oxan-2-yl]oxy}benzoic acid

Step 1

S.N. Materials MW/d Amount/mmol Equiv/Vol 1 Xylose Tributyrate 360 12g/33.3 1.0 (X3B) A 2 Compound B 228 11.4 g/50 1.5 3 Triphenylphosphine262 13.1 g/50 1.5 4 Di-t-butyl 230 11.5 g/50 1.5 azodicarboxylate (DTAD)5 THF 240 mL 20 Vol

Compound A, B and TPP were dissolved in THF and stirred at 0° C. To thismixture was added DTAD and stirring was continued at 0° C. for 1 h, thenat room temperature overnight. The reaction mixture was concentrated.NMR of the crude product showed a mixture of C and D (ratio 1:0.9).Multiple purifications by column chromatography using 0-30% ethylacetate in hexanes provided the desired p isomer C (7 g, 32%) and aisomer D (4.6 g, 21%).

Step 2A:

S.N. Materials MW/d Amount/mmol Equiv/Vol 1 Compound C 570 4.2 g/7.371.0 2 10% Pd/C 200 mg 3 MeOH 50 mL 12 Vol

Compound C was dissolved in methanol and stirred at room temperature. Tothis mixture was added 10% Pd/C. The suspension was stirred under ahydrogen atmosphere at room temperature overnight. The reaction mixturewas filtered through Celite and washed with methanol. The combinedfiltrate and washing were concentrated. The residue was purified by ISCOusing 0-5% MeOH in DCM to give 2.1 g (60%) of pure product 187 and 850mg of impure product.

Step 2B:

S.N. Materials MW/d Amount/mmol Equiv/Vol 1 Compound 4 570 2.8 g/4.9 1.02 10% Pd/C 150 mg 3 MeOH 40 mL 14 Vol

Compound 4 was dissolved in methanol and stirred at room temperature. Tothis mixture was added 10% Pd/C. The suspension was stirred underhydrogen atmosphere at room temperature overnight. The reaction mixturewas filtered through Celite and washed with methanol. The combinedfiltrate and washing were concentrated. The residue was purified by ISCOusing 0-5% MeOH in DCM to give 936 mg (40%) of pure product 186.

Compound 188:5-butanamido-2-{[(2S,3R,4S,5R)-3,4,5-tris(butanoyloxy)oxan-2-yl]oxy}benzoicacid

Compound 183 in metabolic application (50 mg, 0.10 mmol, 1 equiv) wasdissolved in 0.25 mL of DCM, followed by addition of butyric anhydride(0.05 mL, 0.3 mmol, 3 equiv). The reaction was stirred at roomtemperature for 40 minutes, then purified by column chromatography(0-100% EtOAc in hexanes) to yield the title compound as a white solid(42.5 mg, 0.075 mmol, 75% yield). ¹H NMR (400 MHz, DMSO-d₆) δ 12.71 (s,1H), 9.90 (s, 1H), 7.86 (d, J=2.8 Hz, 1H), 7.65 (dd, J=9.0, 2.7 Hz, 1H),7.10 (d, J=9.0 Hz, 1H), 5.36 (d, J=7.2 Hz, 1H), 5.29 (t, J=9.0 Hz, 1H),5.04 (dd, J=9.2, 7.2 Hz, 1H), 4.94 (td, J=9.2, 5.4 Hz, 1H), 4.04 (dd,J=11.5, 5.4 Hz, 1H), 3.70 (dd, J=11.6, 9.4 Hz, 1H), 2.33-2.11 (m, 8H),1.65-1.41 (m, 8H), 0.93-0.77 (m, 12H).

Compound 189:5-amino-2-{[(2S,3R,4S,5R)-3,4,5-tris[(3,3,4,4,4-²H₅)butanoyloxy]oxan-2-yl]oxy}benzoicacid

This compound was prepared as described for compound 183 with theexception that starting materials appropriately enriched in deuterium.LCMS: (M+H+) 511.3. 1H NMR (400 MHz, DMSO-d6) δ 12.41 (s, 1H), 6.86 (d,1H), 6.81 (d, 1H), 6.64 (dd, 1H), 5.27 (t, 1H), 5.14 (d, 1H), 5.00 (dd,1H), 4.92 (td, 1H), 4.02 (dd, 1H), 3.63 (dd, 1H), 2.33-2.14 (m, 6H)

Compound 190:2-{[(2R,3R,4S,5R,6R)-3,4,5-tris(butanoyloxy)-6-[(butanoyloxy)methyl]oxan-2-yl]oxy}benzoicacid

This compound was prepared according to a modified procedure for thepreparation of compound 41.

Compound 191:5-amino-2-{[(2R,3R,4R,5R)-3,4,5-tris[(3,3,4,4,4-²H₅)butanoyloxy]oxan-2-yl]oxy}benzoicacid

This compound was prepared according to a modified procedure for thepreparation of compound 41.

Compound 192:5-amino-2-{[(2S,3R,4S,5R)-3,4-bis[(3,3,4,4,4-²H₅)butanoyloxy]-5-hydroxyoxan-2-yl]oxy}benzoicacid

This compound was prepared according to a modified procedure for thepreparation of compound 32. LCMS: (M−H−) 434.2. ¹H NMR (400 MHz,Chloroform-d) δ 7.46 (d, 1H), 7.04 (d, 1H), 6.85 (dd, 1H), 5.35 (d, 1H),5.15 (dd, 1H), 5.06 (t, 1H), 4.17 (dd, 1H), 3.84 (td, 1H), 3.61 (dd,1H), 2.41 (s, 2H), 2.36 (s, 2H)

Compound 193:(2R,3R)-2-[3,4-bis(butanoyloxy)phenyl]-5,7-bis(butanoyloxy)-3,4-dihydro-2H-1-benzopyran-3-ylbutanoate

A mixture of (2R,3R)-2-(3,4-dihydroxyphenyl)chromane-3,5,7-triol (300mg, 1.03 mmol, 1 eq) and TEA (627.50 mg, 6.20 mmol, 863.13 uL, 6 eq) inTHF (8 mL) was cooled to 0° C. and stirred under N2. Then butanoylchloride (110.12 mg, 1.03 mmol, 107.96 uL, 1.00 eq) was dropped to themixture and then heated to 50° C. and stirred for 10 hours. LCMS showeddesired compound was detected. The reaction mixture was concentratedunder reduced pressure to give a residue. The residue was purified byprep-HPLC (column: Phenomenex Luna C18 200*40 mm*10 um; mobile phase:[water (0.05% HCl)-ACN]; B %: 70%-90%,10 min) to give WX-0637(2R,3R)-2-(3,4-bis(butyryloxy)phenyl)chroman-3,5,7-triyl tributyrate(154 mg, 216.61 umol, 20.96% yield, 90.12% purity) as a white solid.LCMS: (M+H⁺) 641.4 @ 1.476 min; LCMS: (M+Na⁺) 663.3 @ 2.346 min. ¹H NMR(400 MHz, Chloroform-d) δ 7.32 (d, J=2.0 Hz, 1H), 7.30-7.23 (m, 1H),7.19 (d, J=8.4 Hz, 1H), 6.67 (d, J=2.3 Hz, 1H), 6.54 (d, J=2.3 Hz, 1H),5.43-5.37 (m, 1H), 5.12 (s, 1H), 2.97 (dd, J=17.8, 4.5 Hz, 1H), 2.86(dd, J=17.9, 2.3 Hz, 1H), 2.57-2.47 (m, 8H), 2.14 (t, J=7.4 Hz, 2H),1.77 (hd, J=7.4, 2.3 Hz, 8H), 1.44 (h, J=7.3 Hz, 2H), 1.08-0.99 (m,12H), 0.74 (t, J=7.4 Hz, 3H)

Compound 194:(2R,3R)-2-[3,4-bis(propanoyloxy)phenyl]-5,7-bis(propanoyloxy)-3,4-dihydro-2H-1-benzopyran-3-ylpropanoate

A mixture of (2R,3R)-2-(3,4-dihydroxyphenyl)chromane-3,5,7-triol (300mg, 1.03 mmol, 1 eq) and TEA (627.50 mg, 6.20 mmol, 863.13 uL, 6 eq) inTHF (8 mL) was cooled to 0° C. and stirred under N2. Then propanoylchloride (573.76 mg, 6.20 mmol, 573.76 uL, 6 eq) was dropped to themixture and the mixture heated to 50° C. and stirred for 10 hours. LCMSshowed desired compound was detected. The reaction mixture wasconcentrated under reduced pressure to give a residue. The residue waspurified by prep-HPLC (column: Phenomenex Luna C18 200*40 mm*10 um;mobile phase: [water (0.05% HCl)-ACN]; B %: 55%-75%,10 min) to give[2-propanoyloxy-4-[(2R,3R)-3,5,7-tri(propanoyloxy)chroman-2-yl]phenyl]propanoate (151 mg, 258.48 umol, 25.01% yield, 97.67% purity) as a whitesolid. LCMS: (M+H⁺) 571.3 @ 1.342 min; LCMS: (M+Na⁺) 593.3 @2.147 min.¹H NMR (400 MHz, Chloroform-d) δ 7.33 (d, J=2.0 Hz, 1H), 7.30-7.23 (m,1H), 7.19 (d, J=8.4 Hz, 1H), 6.67 (d, J=2.3 Hz, 1H), 6.56 (d, J=2.3 Hz,1H), 5.43-5.37 (m, 1H), 5.12 (s, 1H), 2.97 (dd, J=17.8, 4.5 Hz, 1H),2.86 (dd, J=17.9, 2.3 Hz, 1H), 2.63-2.51 (m, 8H), 2.18 (q, J=7.5 Hz,2H), 1.30-1.20 (m, 12H), 0.95 (t, J=7.5 Hz, 3H).

Compound 195:[(2R,3R)-5,7-di(propanoyloxy)-2-[3,4,5-tri(propanoyloxy)phenyl]chroman-3-yl]3,4,5-tri(propanoyloxy)benzoate

Propionic anhydride (2.78 mL, 21.8 mmol) was added dropwise to a stirredsolution of epigallocatechin gallate (0.5 g, 1.09 mmol) in anhydrouspyridine (2.61 mL, 32.6 mmol) at 0° C. under N₂ atmosphere. Theresulting stirred solution was allowed to come to room temperature andreaction was monitored to completion by LCMS. The solution was dilutedwith 30 mL ethyl acetate and washed with H₂O (30 mL), 1M HCl (30 mL),H2O (30 mL), and saturated NaHCO₃ (30 mL). The organic layer was driedover sodium sulfate, filtered, and concentrated by rotary evaporation.The crude residue was purified by flash chromatography (silica, 10-100%ethyl acetate in hexanes) and fractions were concentrated by rotaryevaporation to yield Compound 197 (0.695 g, 70% yield) as a white solid.¹H NMR (DMSO-d6, 400 MHz): δ 7.54 (s, 2H), 7.38 (s, 2H), 6.79 (m, 1H),6.66 (m, 1H), 5.66 (m, 1H), 5.54 (s, 1H), 3.13-3.17 (m, 1H), 2.96 (d,1H), 2.5-2.65 (m, 16H), 1.0-1.2 (m, 24H)

Compound 196:[(2R,3R)-5,7-bis(4-phenylbutanoyloxy)-2-[3,4,5-tris(4-phenylbutanoyloxy)phenyl]chroman-3-yl]3,4,5-tris(4-phenylbutanoyloxy)benzoate

Step 1:

To a solution of 4-phenylbutanoic acid (3 g, 18.27 mmol) and SOCl₂(10.87 g, 91.35 mmol, 6.63 mL) in dichloromethane (50 mL) is added onedrop of DMF, then the mixture stirred at 20° C. for 5 h. The solvent isremoved in vacuum and toluene (20 mL) added to the mixture. The mixtureis concentrated in vacuo to afford 4-phenylbutanoyl chloride (3.5 g,crude).

Step 2:

To a solution of[(2R,3R)-5,7-dihydroxy-2-(3,4,5-trihydroxyphenyl)chroman-3-yl]3,4,5-trihydroxybenzoate (1 g, 2.18 mmol) and K₂CO₃ (4.52 g, 32.72 mmol)in acetonitrile (100 mL) was added a solution of 4-phenylbutanoylchloride (7.97 g, 43.63 mmol) in acetonitrile (10 mL), then the mixturewas stirred at 20° C. for 10 h. The mixture was filtered, and thefiltrate was concentrated in vacuum. The crude product was purified bysilica gel chromatography (petroleum ether/ethyl acetate=20:1-1:1) toafford compound 198 (2.2 g, 1.28 mmol, 58.7% yield) as a white solid.LC/MS (M+H₃₀ ⁺): 1645.1

Compound 197:[(2R,3R)-5,7-diacetoxy-2-(3,4,5-triacetoxyphenyl)chroman-3-yl]3,4,5-triacetoxybenzoate

Acetic anhydride (6.1 mL) was added dropwise to epigallocatechin gallate(2.0 g) in pyridine (20 mL) at 0° C., and the resulting mixture wasstirred overnight at room temperature. Water was added to the reactionmixture, and the solid was filtered and washed with aq. 1N HCl (10 mL)and heptane (20 mL). The solid was then dissolved in dichloromethane andpassed through a silica gel filter column with dichloromethane as amobile phase to furnish compound 199 (1.0 g, 28%) upon evaporation ofvolatiles. ¹H NMR (CDCl₃): δ 7.6 (s, 2H), 7.2 (s, 2H), 6.75 (s, 1H), 6.6(s, 1H), 5.6 (t, 1H), 5.19 (s, 1H), 2.98-3.02 (m, 2H), 2.18-2.28 (m,24H).

Compound 198:[(2R,3R)-5,7-di(butanoyloxy)-2-[3,4,5-tri(butanoyloxy)phenyl]chroman-3-yl]3,4,5-tri(butanoyloxy)benzoate

Butyryl chloride (6.03 mL) was added to a stirred solution ofepigallocatechin gallate (2.0 g) and pyridine (6.28 mL) indichloromethane (20 mL) over 2 h between −5° C. to 5° C. The resultingmixture was stirred overnight at room temperature. The reaction mixturewas then diluted with dichloromethane (100 mL), washed sequentially withwater (50 mL), 2N HCl (50 mL), saturated sodium bicarbonate (50 mL), andbrine. The organic layer was evaporated in vacuo, and the resultingcrude material was purified by flash chromatography (30% ethylacetate/heptane) to give compound 198 (800 mg, 18%). ¹H NMR (CDCl₃): δ7.6 (s, 2H), 7.22 (s, 2H), 6.78 (s, 1H), 6.6 (s, 1H), 5.62 (t, 1H), 5.18(s, 1H), 2.98-3.02 (m, 2H), 2.4-2.6 (m, 16H), 1.6-1.8 (m, 16H),0.92-1.02 (m, 24H).

Compound 199: 3,4,5-triacetoxybenzoic acid

Acetic anhydride (1.65 mL, 17.6 mmol) was added dropwise to a stirredsolution of gallic acid (0.300 g, 1.76 mmol) in anhydrous pyridine (1.41mL, 17.6 mmol) at 0° C. under N2 atmosphere. The resulting stirredsolution was left to come to room temperature and reaction was monitoredto completion by LCMS. The solution was diluted with 20 mL of ethylacetate and washed with 1M HCl (20 mL) and saturated NaCl (20 mL). Theorganic layer was dried over magnesium sulfate, filtered, andconcentrated by rotary evaporation. The crude residue was purified byflash chromatography (silica, 10-90% acetonitrile in water) andfractions were concentrated by rotary evaporation to yield compound 201(0.259 g, 49.7% yield) as a white solid. ¹H-NMR (DMSO-d₆, 400 MHz): δ13.44 (s, 1H), 7.75 (s, 1H), 2.33 (s, 3H), 2.30 (s, 6H). LC-MS: 319.0(M+Na)⁺

Compound 200: 3,4,5-tris({[2-(1H-indol-3-yl)acetyl]oxy})benzoic acid

This compound was prepared following a modified procedure described forcompound 63. LCMS: (M+H+) 642.2. ¹H NMR (400 MHz, DMSO-d6) δ 11.06 (dd,3H), 7.65 (s, 2H), 7.46 (dd, 3H), 7.37 (dd, 3H), 7.22 (dd, 3H),7.14-7.07 (m, 3H), 7.05-6.97 (m, 3H), 3.71 (s, 4H), 3.43 (s, 2H)

Compound 201: 3,4,5-tris(propanoyloxy)benzoic acid

This compound was prepared following a modified procedure described forcompound 199. LCMS (M−H−) 337.1. ¹H NMR (400 MHz, DMSO-d6) δ 13.43 (s,1H), 7.75 (s, 2H), 2.62 (m, 6H), 1.13 (m, 9H)

Compound 202: 3,4,5-tris({[3-(1H-indol-3-yl)propanoyl]oxy})benzoic acid

This compound was prepared following a modified procedure described forcompound 199.

LCMS: (m+H+) 684.2. ¹H NMR (400 MHz, DMSO-d6) δ 13.48 (s, 1H), 10.85 (d,3H), 7.70 (s, 2H), 7.50 (dd, 3H), 7.38-7.29 (m, 3H), 7.13 (dd, 3H),7.10-7.02 (m, 3H), 6.94 (m, 3H), 3.00 (m, 6H), 2.83 (m, 6H)

Compound 203: 6-oxo-6H-benzo[c]chromene-3,8,9-triyl triacetate

To a mixture of 3,8,9-trihydroxybenzo[c]chromen-6-one (0.3 g, 1.23 mmol,1 eg) and acetyl acetate (501.67 mg, 4.91 mmol, 460.25 uL, 4 eg) in DCM(10 mL) was added triethylamine (TEA) (372.94 mg, 3.69 mmol, 512.98 uL,3 eg). The mixture was stirred at 25° C. for 10 hours. TLC indicated onenew spot was detected. The reaction mixture was quenched by additionwater 10 mL and extracted with EtOAc (10 mL×3). The combined organiclayers were washed with brine 20 mL, dried over Na₂SO₄, filtered andconcentrated under reduced pressure to give the residue. The residue waspurified by column chromatography (SiO₂, Petroleum ether/Ethylacetate=3/1 to 1:1). Compound (8,9-diacetoxy-6-oxo-benzo[c]chromen-3-yl)acetate (0.36 g) was obtained as a gray solid. LCMS: (M+H⁺): 371.0 ¹HNMR (400 MHz, CDCl₃) δ 8.2 (s, 1H), 9.9 m, 2H), 7.1 (m, 2H), 2.3 (s,9H).

Compound 204: [6-oxo-8,9-di(propanoyloxy)benzo[c]chromen-3-yl]propanoate

Propionic anhydride (2.61 mL, 20.4 mmol) was added dropwise to a stirredsolution of urolithin C (0.5 g, 2.04 mmol) in anhydrous pyridine (4.92mL, 61.2 mmol) at 0° C. under N₂ atmosphere. The resulting stirredsolution was allowed to come to room temperature and reaction wasmonitored to completion by LCMS. The solution was diluted with 30 mLethyl acetate and washed with H₂O (30 mL), 1M HCl (30 mL), H₂O (30 mL),and saturated NaHCO₃ (30 mL). The organic layer was dried over sodiumsulfate, filtered, and concentrated by rotary evaporation. The cruderesidue was purified by flash chromatography (silica, 10-100% ethylacetate in hexanes) and fractions were concentrated by rotaryevaporation to yield Compound 204 (0.05 g, 6% yield) as a pink solid. ¹HNMR (DMSO-d6, 400 MHz): 5 8.4 (s, 1H), 8.35 (d, 1H), 8.14 (s, 1H), 7.31(d, 1H), 7.23 (m, 1H), 2.73-2.63 (m, 6H), 1.21-1.14 (m, 9H) ppm

Compound 205: [8,9-di(octanoyloxy)-6-oxo-benzo[c]chromen-3-yl] octanoate

To a solution of 3,8,9-trihydroxybenzo[c]chromen-6-one (0.3 g) inacetonitrile (10 mL) was added K₂CO₃ (0.68 g) followed by octanoylchloride (0.8 g). The resulting mixture was stirred at 50° C. for 24hours. Additional octanoyl chloride (0.8 g) was added and the mixturewas stirred at 50° C. for 12 hours. The reaction mixture was quenched byaddition of water (10 mL) and extracted three times with ethyl acetate(10 mL). The combined organic layers were dried over Na₂SO₄, filteredand concentrated under reduced pressure. The residue was purified bycolumn chromatography (SiO₂, petroleum ether/ethyl acetate, 9:1 to 1:1)to give [8,9-di(octanoyloxy)-6-oxo-benzo[c]chromen-3-yl]octanoate (0.45g, 55.5%) as a white solid. ¹H NMR (400 MHz, CDCl₃): δ 8.186 (s, 1H),7.926 (d, 1H), 7.908 (s, 1H), 7.157-7.096 (m, 2H), 2.621-2.573 (m, 6H),1.79-1.75 (6H, m), 1.5-1.25 (m, 24H), 0.916-0.878 (m, 9H) ppm

Compound 206:8,9-bis({[(3R)-3-(butanoyloxy)butanoyl]oxy})-6-oxo-6H-benzo[c]chromen-3-yl(3R)-3-(butanoyloxy)butanoate

This compound was prepared following a modified procedure described forcompound 203.

Compound 207: 8,9-bis(decanoyloxy)-6-oxo-6H-benzo[c]chromen-3-yldecanoate

This compound was prepared following a modified procedure described forcompound 203.

Compound 208:8,9-bis({[3-(1H-indol-3-yl)propanoyl]oxy})-6-oxo-6H-benzo[c]chromen-3-yl3-(1H-indol-3-yl)propanoate

This compound was prepared following a modified procedure described forcompound 63.

Compound 209:8,9-bis({[2-(1H-indol-3-yl)acetyl]oxy})-6-oxo-6H-benzo[c]chromen-3-yl2-(1H-indol-3-yl)acetate

This compound was prepared following a modified procedure described forcompound 63.

Compound 210: 8,9-bis(butanoyloxy)-6-oxo-6H-benzo[c]chromen-3-ylbutanoate

This compound was prepared following a modified procedure described forcompound 203.

Compound 211:(1R)-1-[(2R,3R,4S)-3,4-bis({[3-(1H-indol-3-yl)propanoyl]oxy})oxolan-2-yl]-2-{[3-(1H-indol-3-yl)propanoyl]oxy}ethyl3-(1H-indol-3-yl)propanoate

This compound was prepared following a modified procedure described forcompound 63.

Compound 212:(1R)-1-[(2R,3R,4S)-3,4-bis({[2-(1H-indol-3-yl)acetyl]oxy})oxolan-2-yl]-2-{[2-(1H-indol-3-yl)acetyl]oxy}ethyl2-(1H-indol-3-yl)acetate

This compound was prepared following a modified procedure described forcompound 63.

Compounds 213:(1R)-1-[(2R,3R,4S)-3,4-bis(butanoyloxy)oxolan-2-yl]-2-(butanoyloxy)ethylbutanoate

This compound was prepared following a modified procedure described forcompound 203.

Compound 214:(1R)-2-(acetyloxy)-1-[(2R,3R,4S)-3,4-bis(acetyloxy)oxolan-2-yl]ethylacetate

This compound was prepared following a modified procedure described forcompound 203.

Compound 215: (3R,4S,5R)-3,4,5-triacetoxycyclohex-1-ene-1-carboxylicacid

Acetic anhydride (1.61 mL, 17.2 mmol) was added dropwise to a stirredsolution of shikimic acid (0.300 g, 1.72 mmol) in anhydrous pyridine(1.38 mL, 17.2 mmol) at 0° C. under N2 atmosphere. The resulting stirredsolution was left to come to room temperature and reaction was monitoredto completion by LCMS. The solution was diluted with 20 mL of ethylacetate and washed with 1M HCl (20 mL) and saturated NaCl (20 mL). Theorganic layer was dried over magnesium sulfate, filtered, andconcentrated by rotary evaporation. The crude residue was purified byflash chromatography (silica, 10-90% acetonitrile in water) andfractions were concentrated by rotary evaporation to yield compound 43(0.18 g, 34.8% yield) as a white solid. ¹H-NMR (DMSO-d₆, 400 MHz): δ12.90 (s, 1H), 6.61 (dt, 1H), 5.60 (m, 1H), 5.18 (dd, 1H), 5.12 (dt,1H), 2.76 (m, 1H), 2.35 (m, 1H), 2.04 (s, 5H), 2.01 (s, 3H). LC-MS:299.1 (M−H)⁻

Compound 216:(3R,4S,5R)-3,4,5-tris({[3-(1H-indol-3-yl)propanoyl]oxy})cyclohex-1-ene-1-carboxylicacid

This compound was prepared following a modified procedure described forcompound 63.

Compound 217:(3R,4S,5R)-3,4,5-tris(propanoyloxy)cyclohex-1-ene-1-carboxylic acid

This compound was prepared following a modified procedure described forcompound 215.

Compound 218:(3R,4S,5R)-3,4,5-tris(butanoyloxy)cyclohex-1-ene-1-carboxylic acid

This compound was prepared following a modified procedure described forcompound 215.

Compound 219: (2R,3R,4S,5S)-tetrahydro-2H-pyran-2,3,4,5-tetrayltetraacetate

L-Arabinose (50 g), N,N-dimethylpyridin-4-amine (6 g), and triethylamine(367 mL) were dissolved in 700 mL DCM and stirred at 0° C. under N₂.Acetic anhydride (217 mL) was added dropwise over 30 minutes and thereaction mixture was stirred overnight. The solvent was removed underreduced pressure and the residue was redissolved in ethyl acetate,washed with 1M HCl, H₂O and brine, dried over MgSO₄ and evaporated.Purification on normal phase with 0-100% ethyl acetate in hexanes gaveCompound 53 as a waxy/amorphous solid (50% yield). 1H-NMR (DMSO-CDCl₃,400 MHz): δ 6.35 (d, 1H), 5.37 (m, 3H), 4.06 (dd, 1H), 3.82 (dd, 1H),2.155 (s, 3H) 2.15 (s, 3H), 2.02 (s, 6H). LC-MS: 341.1 (M+Na)⁺

Example 2. In Vitro Assays

Acylated active agents disclosed herein may be stable under a range ofphysiological pH levels and cleaved selectively at a desired site ofaction (for example, in the GI tract, e.g., in the stomach, smallintestine, or large intestine) by enzymes present in the localmicroenvironment. Acylated active agents are tested for chemicalstability at a range of pH levels as well as their ability to bedegraded in representative in vitro systems. Data for select acylatedactive agents are shown below.

Assay 1. Stability of acylated active agents in Simulated Gastric Fluid(SGF). This assay was used to assess the stability of an acylated activeagent in a stomach.

Medium was prepared by dissolving 2 g of sodium chloride in 0.6 L inultrapure water (MilliQ®, Millipore Sigma, Darmstadt, Germany). The pHwas adjusted to 1.6 with 1N hydrochloric acid, and the volume was thenadjusted to 1 L with purified water.

60 mg FaSSIF powder (Biorelevant™, London, UK) were dissolved in 500 mLbuffer (above). Pepsin was added (0.1 mg/mL) (Millipore Sigma,Darmstadt, Germany), and the solution was stirred. The resulting SGFmedia were used fresh for each experiment.

Test compounds were dissolved in DMSO stock to 1 mM. An aliquot of theDMSO stock solution was removed and diluted in the SGF Media in 15 mLfalcon tubes to generate a total compound concentration of 1 μM. A 1 mLaliquot was immediately removed and diluted once with 1 volume ofacetonitrile for T0 timepoint. The mixture was sealed and mixed at 37°C. in an incubator. Aliquots (1 mL) were removed at regular intervalsand immediately quenched by the addition of 1 volume of acetonitrile.The resulting samples were analyzed by LC/MS to determine degradationrates in SGF.

Assay 2. Stability of acylated active agents in Simulated IntestinalFluid (SIF). This assay was used to assess the stability of an acylatedactive agent in a small intestine.

Phosphate buffer was prepared by dissolving 0.42 g of sodium hydroxidepellets and 3.95 g of monobasic sodium phosphate monohydrate and 6.19 gof sodium chloride in ultrapure water (MilliQ®, Millipore Sigma,Darmstadt, Germany). The pH was adjusted to 6.7 using aq. HCl and aq.NaOH, as necessary, and the solution was diluted with ultrapure water toproduce 1 L of the pH 6.7 buffer.

112 mg FaSSIF powder (Biorelevant™, London, UK) was dissolved in 50 mLof the pH 6.7 buffer. 2 to 3 mL of the resulting solution were thenadded to 500 mg pancreatin (Millipore Sigma, Darmstadt, Germany). Theresulting mixture was agitated by finger tapping the vessel containingthe mixture until milky suspension formed. At this time, the remainderof the 50 mL FaSSiF/pH 6.7 buffer solution was added. The resultingsuspension was flipped upside down 10 times to produce SIF, which wasused fresh.

Test compounds were dissolved in DMSO stock to 1 mM. An aliquot of theDMSO stock solution was removed and diluted in the SIF media in 15 mLfalcon tubes to produce a mixture with a tested compound concentrationof 1 μM. A 1 mL aliquot was immediately removed and diluted once with 1volume of acetonitrile for T0 timepoint. The mixture was sealed andagitated at 37° C. in an incubator. Aliquots (1 mL) were removed atregular intervals and immediately quenched by the addition of 1 volumeof acetonitrile. The resulting samples were analyzed by LC/MS todetermine degradation rates.

Assay 3. In vitro Colonic Material Stability Assay. This assay was usedto assess the stability of an acylated active agent in a largeintestine. All experiments were performed in an anaerobic chambercontaining 90% nitrogen, 5% hydrogen and 5% carbon dioxide. Colonicmaterial was resuspended as a slurry (15% w/v final concentration) inpre-reduced, anaerobically sterilized dilution blanks (Anaerobe SystemsAS-908). The colonic material was then inoculated into 96 well platescontaining YCFAC media (Anaerobe Systems AS-680) or other suitable media(6.7 μL slurry into 1 mL total media). Compounds or groups of compoundswere added to each individual well to reach a final analyteconcentration of 1 or 10 μM, and the material was mixed by pipetting.Sample was removed after set timepoints (0, 120, 240, 480, 1440, 2880minutes after initiation of the assay), quenched with acetonitrilecontaining internal standard, and analyzed by LC/MS.

Buffer Assay. Stability of acylated active agents in a buffer. Thisassay provides for the assessment of the stability of an acylated activeagent at different physiological pH levels.

Compounds are diluted in DMSO, and added in the appropriate quantity tophosphate buffer (pH levels 2, 4, 6, and 8) to reach a total sampleconcentration of 2 μM. Compounds are incubated at RT, and aliquots areremoved at time points 0, 60, 120, 360 and 1440 minutes and analyzed forpurity by LC/MS/MS.

TABLE 1 Assay 1 (SGF) Assay 2 (SIF) Assay 3 (% Remaining (% @ Remaining(% Remaining Compound @ 1 hour) 4 hours) at 24 h) 1 C B 2 C A A 4 C A 5B 8 C A 9 C A 10 B A 11 C 12 C A 13 B A 14 C A C 16 C C 17 A 18 C B C 19C B B 21 C C B 22 C B B 23 B C 25 C C 26 C A 27 C B 28 C 29 C C 31 C A32 B C 36 C C 38 C C 39 C B C 40 C C C 41 C C 42 C 45 A A 46 C A A 47 CA A 48 49 Run 1: B, A A Run 2: C 50 C A A 51 C C B 52 B B B 53 C B C 54B A A 55 C A A 56 A A 57 C A A 58 A A 59 A A 60 A A 61 A A 62 C A C 64 AA 68 C A B 69 B A A 70 B A Run 1: A, Run 2: B 71 B A A 72 C B A 73 C A A74 C A A 75 C 76 C A A 77 C C A 78 B A A 80 C A A 81 B A 83 C A A 84 C AA 85 B A A 86 B A 87 B A 88 A A 90 C B A 91 C A A 92 C A A 93 C A A 94 CA A 95 B 96 C A A 97 B A 98 B A A 99 B A 100 B A 101 C A A 102 C B A 103C A A 104 C A A 105 C B B 106 C A A 107 B A B 108 C A A 109 C A A 110 CA 111 C A A 112 C A A 113 C A C 114 C A A 115 C A A 116 C 117 C A A 118C A 119 A 120 A 121 A 122 A 123 Batch 1: C; C A Batch 2: B 124 C A 125 CA 126 C A A 127 C B 128 C 129 C A 130 C A 131 A 132 C 136 C A 141 C 147A 149 B A A 150 A A 151 C A 158 A A 159 C 161 C C B 162 C A C 163 C C B165 B A A 166 B A A 167 B A A 168 B A A 169 C C 170 B A A 171 C A 172 CA B 173 C A 174 C 175 A 176 A 177 C 181 C A 182 A 183 B B 184 B 185 C BB 186 C A A 188 A 189 B 192 B 193 C A A 194 B A A 195 C 197 A 198 Runs 1and 2: C; Run 3: B 199 C B 200 C A A 201 C A A 202 A C 203 B 204 C A 208B B 210 A 215 B 216 B B 217 B 218 B 219 C A AIn Table 1, A: <25% of the tested compound remaining; B: 25-75% of thetested compound remaining; and C: >75% of the tested compound remaining.

Table 1 shows that, for example, compounds 2, 4, 8-10, 12-14, 18, 19,22, 26, 27, 31, 39, and 221 can be selectively delivered to the upperintestine.

Example 3. In Vivo Evaluation of an Acylated Active Agent

Acylated active agents disclosed herein may be useful in modulatingautoimmunity markers and for treating autoimmune disorders. This exampledemonstrates the capability of an exemplary acylated active agent,compound 2, to induce CD4⁺CD25⁺ Treg cells (an autoimmunity marker) in asubject.

C57BL/6 mice were divided into seven cohorts, as listed in Table 2.

TABLE 2 # of Model Treatment* animals Dose** Frequency Route HFD-fed ND10 Ad libitum Diet C57BL/6 HFD 10 Ad libitum Diet mice HFD + Acetate 105% Ad libitum Diet HFD + EGCG 10 1% Ad libitum Diet HFD + Acetate + 105% + 1% Ad libitum Diet EGCG Compound 2 10 6% Ad libitum Diet (EGCG-8A)HFD + 10 0.45 mg/g Ad libitum Diet rosiglitazone *In Table 2, ND meansnormal diet, HFD means high-fat diet, and EGCG means epigallocatechingallate. **In Table 2, dose percentages refer to weight percentagerelative to the high fat diet. The results of this study are illustratedin the FIG. 1, which shows a synergistic anti-inflammatory effect ofCompound 2 in animals fed a high-fat diet, as compared to theadministration of EGCG, acetate, or their combination as separatecompounds.

Example 4. Impact of Compounds on Inflammatory Signals in Primary HumanPeripheral Blood Mononuclear Cells (PBMCs)

Human donor blood (8 mL) was collected in sodium citrate CPT tubes andcentrifuged at 1,600×g for 20 minutes at room temperature. Buffy coatcontaining PBMCs was collected and transferred to a 50 mL conical tubecontaining 30 mL of RPMI-1640 medium at room temperature (supplementedwith penicillin-streptomycin). PBMCs samples were centrifuged at 400×gfor 10 minutes at 10° C. The pelleted PBMCs were washed twice in 10 mlof RPMI-1640 medium (supplemented with penicillin-streptomycin), thenresuspended in RPMI-1640 medium (supplemented withpenicillin-streptomycin, fetal bovine serum, and L-Glutamine). PBMCswere filtered through a 70 micron mesh to remove any cellular debris.The volume was adjusted to achieve 1.66×10⁶ cells/mL, from which 180 μl(300,000 PBMCs) were added into each well in a 96-well plate (sterile,tissue culture treated, round bottom). PBMCs in a 96-well plate wererested for 30 minutes in a 37° C. 5% CO₂ incubator, then subsequentlytreated with 10 μl of indicated compound. After 2 hours 10 μL of LPS(O111:B4) 1 mg/mL was added to test wells. After 24 hours of incubationat 37° C. 5% CO₂, 100 μL of cell supernatant was collected andtransferred to a 96-well plate (non-tissue treated, flat bottom). Theplate was centrifuged at 350×g for 5 minutes at room temperature, andthen the clear supernatant transferred to a new 96-well plate(non-tissue treated, flat bottom). The remaining cells were tested forviability using CellTiter-Glo® Luminescent Cell Viability Assay(Promega). The supernatant was analyzed for TNFα, IL-6 and IL-1/? (kitLXSAHM-03; R&D Systems), using Luminex Immunoassay Technology (MAGPIXSystem). Cytokine levels of LPS treated DMSO control samples were set to100%, and compound treated samples were expressed relative to this(Table 3).

TABLE 3 TNFα IL6 IL1β Concentration % of DMSO % DMSO % DMSO Compound(uM) control control control Propionate 100 + + + Arabinose 100 + + =butanediol 100 = = = Beta- 100 − = − hydroxybutyrate (BHB) Butyrate 100++ + − Acetate 100 = = = Quercitin 100 + + + (−) >110% DMSO; (=) 90% ><110% DMSO (+) 50% > <90% DMSO (++) <50% DMSO

These data demonstrate that acylated active agents (e.g., thoseincluding propionate, butyrate, arabinose, and/or quercetin) canmodulate autoimmunity markers (e.g., reduce TNFα, IL6, and/or IL1βlevels).

Example 5. In Vivo Assessment of Compounds in DSS Colitis

Specific pathogen-free female Swiss Webster mice, 6-8 weeks old,weighing 16-20 g were obtained from Harlan. Mice were randomized intotreatment groups (n=10) based on body weight on study day −2. From studyday 0 through 5, all groups were given 3% dextran sulfate sodium (DSS,Spectrum, Lot #2DC0020) in drinking water. On study day 5 through,drinking water was switched to water without DSS until the study end onday 7. From study day 0 through 7, the negative control (vehicle),positive control (cyclosporine A), and the treatment groups were givenvehicle (1% Methylcellulose (Sigma, 400 cP)), cyclosporine A (70 mg/kg),and compound 44 (100 mg/kg) once a day until study day 7 via oralgavage. On study day 7, animals were anesthetized with Isoflurane andbled to exsanguination followed by cervical dislocation. The entirecolon was removed and the overall efficacy of compound 44 was assessedbased on colon histopathology. At the end of the study, colonhistopathology showed significant reduction in mucosal thickness bycompound 44 compared to the vehicle control (FIG. 2). Statisticalanalysis was performed with GraphPad Prism (GraphPad Software).Mann-Whitney U test was used to assess significance between the vehiclecontrol and each treatment group.

This data is relevant to immune mediated IBD, driven primarily though Tcell biology. A positive result in this pharmacology in vivo studysuggests efficacy in human IBD.

Example 6: Adoptive T-Cell Transfer Study

On study day 0, naïve T-cells were obtained from the spleen of 11 to 12week-old Balb/C mice using the SCID Cell Separation Protocol with thefollowing labelling scheme; CD4+CD45RB^(high)CD25− andCD4+CD45RB^(low)CD25+ cell (APC CD4 Antibody (100412), FITC CD45RBAntibody (103306), PE CD25 Antibody (101904), from BioLegend, San Diego,Calif., and CD4 cell enrichment kit: 19752A, Stemcell Technologies,Cambridge, Mass.) and the 4×10⁶ of CD4+CD45RB^(high)CD25− cells weretransferred 6-7 weeks old CB-17 SCID mice. Recipient mice wererandomized based on their body weight (n=12 mice/group), and received200 ml of vehicle (0.5% methyl-cellulose) (vehicle control), 0.5 mg permouse of anti-IL-12 (positive control), 30 mg/kg of olsalazine (positivetreatment group), and 100 mg/kg of compound 44 (compound treatmentgroup) through day 42. On day 42, animals were anesthetized withIsoflurane and bled to exsanguination followed by cervical dislocation.The entire colon was removed and the overall efficacy of compound 44 wasassessed based on colon length and weight per length. At the end of thestudy, colon length (FIG. 3A) and weight per length (FIG. 3B) weresignificant increased and reduced, respectively by compound 44 comparedto the vehicle control, whereas olsalazine significantly increased thecolon weight per length, but not the colon length. Statistical analysiswas performed with GraphPad Prism (GraphPad Software). Mann-Whitney Utest was used to assess significance between the vehicle control andeach treatment group.

This data is relevant to immune mediated IBD, driven primarily though Tcell biology. A positive result in this pharmacology in vivo studysuggests efficacy in human IBD.

Example 7: Neutrophil Chemokine Production Assay

A volume 25 mL of human blood was layered over 15 mL of Histopaque®-1077and centrifuged at 500 g, RT, for 30 min with no break applied to thecentrifuge. The PBMC band and Histopaque®-1077 layer were removedleaving behind the bottom red layer which was mixed with 40 mL of 1× redblood cell (RBC) lysis buffer (Sigma-Aldrich) was and split into two 50mL tubes. The volume for both fractions was brought to 50 mL with RBClysis buffer, mixed by inversion, and then incubated at RT for 10 min.Solutions were centrifuged at 250 g, for 10 min at RT and thesupernatant liquids removed. The reddish pellets were re-suspended in 1mL of RBC lysis buffer and combined. The cell suspension was incubatedfor 5 min at RT in RBC lysis buffer. After incubation 45 mL of HanksBalanced Salt Solution with no calcium, magnesium or phenol red (HBSS−)was added, the cell suspension was spun (250 g for 10 min at RT), andsupernatant liquids were removed. The white pellet was re-suspended in 1mL of HBSS−, and cell counts were determined. The neutrophil cellsuspension was brought to a concentration of 1.11e6 cells/mL in RPMIcomplete (Sigma-Aldrich), and 180 μL of cell suspension was transferredto all wells within a sterile 96-well tissue culture treated plateresulting in 2.0e5 cells/well. Test compounds were brought to a 20×concentration in RPMI with 2% DMSO, and 10 μL of compound solutions wereadded to wells respective for each compound and incubated for 30 min.After incubation, 10 μL of 2 μg/mL LPS solution in RPMI complete wasadded to each well except for control wells, which received anadditional 10 μL of media. Cells were incubated for 12 h (37° C. 5%CO₂), after which plates were centrifuged at 250 g, RT, for 5 min andsupernatant liquids were obtained and stored at −80° C. until analyzedvia Luminex® Multiplex Assay for various chemokines and cytokines. Threedata points were acquired from two different blood donors and averaged.Statistical analysis was performed using a two-tailed t-test comparingchemokine/cytokines production in the presence of each individualcompound to the DMSO+LPS positive control.

TABLE 4 % IL-8 % MIP-1α % MIP-1β (Vehicle + (Vehicle + (DMSO + Conc. LPS= LPS = LPS = Compound (μM) 100%) 100%) 100%) acetate 500.0 − + +acetate 1000.0 − ++ + acetate 3000.0 +++ +++ +++ L-arabinose 500.0 − − −L-arabinose 1000.0 − − − EGCG 0.1 − − − EGCG 1.0 − − − quercetin 0.1 − −− quercetin 1.0 − − − butanediol 100.0 − − + butanediol 500.0 − − +beta- 200.0 − − + hydroxybutyric acid beta- 2000.0 + ++ + hydroxybutyricacid butyrate 500.0 ++ +++ +++ butyrate 1000.0 ++ +++ +++ propionate500.0 − +++ +++ propionate 1000.0 ++ +++ +++ propionate 3000.0 +++ ++++++ Vehicle + LPS = 100% − = >90% Vehicle + = <90% Vehicle ++ = <70%Vehicle +++ = <50% Vehicle

Neutrophils are the first response from the innate immune system. Thereis a clear link between neutrophil presence and disease activity inulcerative colitis. IL-8/MIP1a and b are important CC chemokinesproduced from neutrophils. Compounds indicated in Table 4 reduceneutrophil production of these mediators and, therefore, may be usefulin a variety of autoimmune disorders, e.g., inflammatory bowel disease,rheumatoid arthritis, and scleroses.

Example 8: Investigating AhR Activation in Caco-2 Cells Through CYP1A1mRNA Expression

Caco-2 cells from American Type Culture Collection (ATCC) were plated ina sterile tissue culture treated 96-well plate (ThermoFisher) at 8.0×10⁵cells per well, and grown overnight at 37° C. 5% CO₂ in DMEM complete(Gibco) in order to achieve confluence. After the incubation medium wasaspirated off of the Caco-2 monolayers, tissues were then washed with200 μL of warmed PBS solution, and subsequently 190 μL of pre-warmedgrowth medium was added to each well. Compounds of interest were dilutedat a 20× concentration in growth medium containing 2% DMSO, and 10 μL ofcompound solutions were added to respective wells in triplicate.Compounds where incubated overnight at 37° C. 5% CO₂. 2-(1¹H-indole-3′-carbonyl)-thiazole-4-carboxylic acid methyl ester (ITE) wasused as the positive control for AhR activation at 1 and 100 μMconcentrations. At the end of the incubation, medium was aspirated offof the Caco-2 cells, and the cells washed with 100 μL of cold PBSsolution. RNA was extracted via the TaqMan™ Gene Expression Cells-to-CT™Kit (ThermoFisher) according to the manufacturers protocol. TheQuantStudio 6 Flex (Applied Biosciences) was used to analyze mRNA levelsof CYP1A1 using GAPDH as the endogenous control. TaqMan™ probe sets forboth genes were acquired from ThermoFisher. Samples were run intriplicate and data was analyzed using the QuantStudio software andreported as linear (Table 1) and log 2(ΔΔC_(T)) values. Statisticalanalysis was performed using a two-tailed t-test comparing CYP1A1 levelsin the presence of each individual compound to the vehicle negativecontrol.

Activation of AHR has been with associated with immune modulation andactive compounds (+, ++, +++) may be beneficial in treating a variety ofinflammatory and autoimmune diseases including ulcerative colitis,multiple sclerosis, rheumatoid arthritis.

TABLE 5 Conc. Average CYP1A1 (μM) mRNA levels vehicle control N/A −acetate 1000.0 − acetate 3000.0 − L-arabinose 1000.0 − EGCG 0.1 − EGCG1.0 − quercetin 0.1 − quercetin 1.0 + butanediol 500.0 −beta-hydroxybutyric acid 2000.0 − butyrate 1000.0 − butyrate 3000.0 −propionate 1000.0 − propionate 3000.0 − Indole-3-acetic acid 500.0 −Indole-3-acetic acid 1000.0 − Indole-3-butyric acid 500.0 −Indole-3-butyric acid 1000.0 − Indole-3-propionic acid 500.0 −Indole-3-propionic acid 1000.0 − indole 1000.0 + Indole-3-aldehyde1000.0 + indole-3-carbinol 1000.0 + Indole-3-acetic acid 500.0 +++Indole-3-acetic acid 1000.0 ++ Indole-3-carboxylic acid 1000.0 −Indole-3-acrylic acid 10.0 +++ Indole-3-acrylic acid 100.0 +++Indole-3-acrylic acid 1000.0 +++ Indole-3-pyruvic acid 10.0 +++Indole-3-pyruvic acid 100.0 +++ Indole-3-pyruvic acid 1000.0 +++ ITE 1μM 1.0 +++ Urolithin B 10.0 ++ Urolithin B 100.0 +++ Urolithin C 10.0 ++Urolithin C 100.0 ++ 4-hydroxy-3-methylbenzoic acid 10.0 +++4-hydroxy-3-methylbenzoic acid 100.0 +++ Benzoic acid 10.0 + Benzoicacid 100.0 + Hydrocinnamic acid 10.0 + Hydrocinnamic acid 100.0 +L-tryptophan 10.0 ++ L-tryptophan 100.0 +++ D-tryptophan 10.0 ++D-tryptophan 100.0 +++ L-homoserine 5.0 + L-homoserine 50.0 + L-arginine5.0 + L-arginine 50.0 + Myricetin 10.0 + Myricetin 100.0 ++Indole-3-lactic acid 10.0 + Indole-3-lactic acid 100.0 +++4-hydroxyphenylpyruvic acid 10.0 ++ 4-hydroxyphenylpyruvic acid 100.0+++ Pterostilbene 10.0 ++ Pterostilbene 100.0 ++ Astaxanthine 10.0 +Astaxanthine 100.0 + 3,4-dihydroxyphenylacetic acid 10.0 +3,4-dihydroxyphenylacetic acid 100.0 ++++ δ-tocopherol 10.0 +δ-tocopherol 100.0 +++ 1-methylindole-3-alanine 10.0 +1-methylindole-3-alanine 100.0 +++ Piceatannol 10.0 + Piceatannol 100.0+++ Kynurenine 10.0 + Kynurenine 100.0 ++ In Table 5, vehicle =baseline; − = <2-fold Vehicle; + = >2-fold Vehicle; ++ = >5-foldVehicle; +++ = >10-fold Vehicle; ++++ = >40-fold Vehicle

Example 9: Human Caco-2 Barrier Integrity Assay

Caco-2 colonocytes were maintained at 37° C. and 5% CO₂ in Dulbecco'sModified Eagle Medium (DMEM) and supplemented with 10% FBS, 1% NEAA, and1% penicillin-streptomycin. At 70-80% confluency, cells were trypsinizedand seeded in 0.4 cm² transwell collagen I coated membranes withsupplemented DMEM in both apical and basolateral compartments. Cellswere seeded at a density of 200,000 cells per well and maintained for 10days to form a polarized barrier with a TransEpithelial ElectrialResistance (TEER) reading above 1000Ω. On the first day of the assay,initial TEER readings were taken and cytokines were added to thebasolateral media (50 ng/mL TNFα, 25 ng/mL IFNγ and 10 ng/mL IL-1β) toreduce barrier integrity while compounds diluted in (dimethyl-sulfoxide)DMSO were added to the apical media in triplicate. After 48 hours, TEERreadings were taken again and viability was measured by CellTiter 96®AQueous One Solution Cell Proliferation Assay (Promega). The percentchange in TEER over the 48 hours was determined and normalized to the0.1% DMSO control (Table 6). None of the compounds reduced proliferationand therefore did not alter cell viability.

TABLE 6 % Change in TEER from DMSO No treatment ++ (170%) DMSO +cytokines − (100%) Acetate 1 mM + cytokines − Acetate 3 mM + cytokines −Arabinose 0.5 mM + cytokines − Arabinose 1 mM + cytokines − EGCG 100nM + cytokines − EGCG 1 μM + cytokines − Quercetin 100 nM + cytokines −Quercetin 1 μM + cytokines + Butanediol 100 μM + cytokines − Butanediol0.5 mM + cytokines − BHB 200 μM + cytokines + BHB 2 mM + cytokines −Butyrate 1 mM + cytokines − Butyrate 3 mM + cytokines − Butyrate 5 mM +cytokines ++ Propionate 1 mM + cytokines + Propionate 3 mM + cytokines++ Statistical changes in TEER were determined by way ANOVA and comparedto DMSO. <125%: − 125% > <150%: + 150% > <200%: ++ 200% >: +++

Barrier function and integrity is an important feature of a variety ofdiseases and can be a hallmark of a damaged GI tract. Inflammation candrive a reduction of barrier function. By improving TEER lesstranslocation of bacteria and bacterial products occur, thus dampeningthe immune response and damage to the GI tract and systemic immunesystem. This is important for the following disease areas: leaky gut,IBD/autoimmune, metabolic disorders, NASH, multiple sclerosis.

Example 10: Human Regulatory T Cell Differentiation Assay

Peripheral blood mononuclear cells (PBMCs) from whole blood donated byhealth volunteers were separated by Ficoll-Paque gradient centrifugationand naïve CD4⁺ T cells were subsequently isolated using magnet beads(EasySep™ Human Naïve CD4⁺ T Cell Isolation Kit, Cambridge, Mass.). Forregulatory T cell (Treg) differentiation assay, naïve CD4⁺ T cells werecultured (1-10×10⁴ cells) in CTS OpTmizer medium for 6 days andstimulated with 5 ng/ml TGF-β, 100 U/ml IL-2, and ImmunoCult™ HumanCC3/CD28/CD2 T Cell Activator; Stemcell #10990) with/without ourCompounds. Cell viability was determined using a viability dye(eBioscience Fixable Viability Dye eFluor 780: ThermoFisher65-0865-14)at 1:500 dilution. The cells were gated for Treg, defined as Live,CD11c, CD14⁻, CD19⁻, CD8, CD4⁺, CD3⁺, CD25⁺, FOXP3⁺. Percent (%) Tregswere calculated as percentage of CD4⁺, CD25⁺, FOXP3⁺ cells over totalCD4⁺ T cells. Statistical analysis was performed with GraphPad PrismSoftware Using One-Way ANOVA.

TABLE 7 Treg induction Cell viability Treatment % DMSO % DMSO Aceticacid 1 mM + = Acetic acid 3 mM ++ − L-Arabinose 0.5 mM = = L-Arabinose 1mM = = EGCG 100 nM = = EGCG 1 uM = = Quercetin 100 nM − − Quercetin 1 uM= = (R)-1,3-Butanediol 100 uM = = (R)-1,3-Butanediol 0.5 mM − − SodiumBHB 2 mM + = Sodium BHB 20 mM − − Butyric Acid 3 mM − − Propionic acid 3mM ++ = Rosiglitazone 10 uM = = Rosiglitazone 100 uM = − Obeticholicacid 100 uM + = DMSO = (100.0) = (100%) <90%: − 90% > <110%: = 110% ><130%: + 130% >: ++

Compounds that increased the differentiation of naïve CD4⁺ T cells intoTregs are indicated as (+) or (++). Compounds that decreased thedifferentiation of naïve CD4⁺ T cells into Tregs are indicated as (−).Tregs play an important role in keeping the balance of the immunesystem, and compounds that increase Tregs (+, ++) may be useful in thetreatment of autoimmune and inflammatory diseases, whereas compoundsthat reduce Tregs (−) may enhance the efficacy of immunotherapy incancer patients.

These data demonstrate that acylated active agents (e.g., thoseincluding) can modulate autoimmunity markers (e.g., increase Treg cellcounts).

Example 11: Ulcerative Colitis Animal PK Studies

Detection of D5-Butyrate in Feces after Dosing with Compound 189

Three female CD-1 mice, single housed in separate metabolism cages, werefasted for 2-4 hours before administration via oral gavage of a 10 mg/mLsuspension of compound 189 in 1% methyl cellulose at a dose volume of 10mL/kg. Feces were collected at the following intervals (0-4 hrs, 4-6hrs, 6-8 hrs, 8-12 hrs and 12-24 hrs). Samples were frozen and stored at−80° C. until analysis. Quantification of butyrate on the samples wasdone via a modification of J. Han et at Analytica Chimica Acta 854(2015) 86-94.

Detection of Mesalamine and Compound 44 in Feces after Dosing withCompound 44

Five female Swiss Webster mice were placed in metabolic cages and wereadministered via oral gavage of a 10 mg/mL suspension of compound 44 in1% methyl cellulose at a dose volume of 10 mL/kg. Feces were collectedat the following intervals (0-4 hrs, 4-8 hrs, 8-24 hrs). Samples werefrozen and stored at −70° C. until analysis. Quantification ofmesalamine and compound 44 were done via bioanalysis using LC-MS/MS.

Detection of Compound 44 in Whole Blood after Dosing with Compound 44

Nine female Swiss Webster mice were placed individually housed cages andwere administered via oral gavage of a 10 mg/mL suspension of compound44 in 1% methyl cellulose at a dose volume of 10 mL/kg. Whole blood (125uL) was collected submandibular and transferred to chilled K₂EDTA tubespre-filled with 125 uL of acetonitrile and frozen at −70° C. untilanalysis. Mice (1-3) were sampled at 5 min, 1 hr, and 8 hr. Mice (4-6)were sampled at 15 min, 2 hours and 24 hours. Mice (7-9) were sampled at30 min and 4 hrs. Quantification of compound 44 was done via bioanalysisusing LC-MS/MS.

The results of the studies are shown in Table 8 below.

TABLE 8 Compound Medium Concentration, uM Mesalamine¹ Feces 224.18Compound 44¹ Feces 10.72 Compound 44¹ Blood 0.68 d5-Butyrate² Feces397.25 ¹Detected after dosing with Compound 44 ²Detected after dosingwith Compound 189

Example 12: In Vivo Rheumatoid Arthritis Model

A study was conducted to determine the efficacy of test articles dosedby the oral (PO) route for inhibition of the inflammation (pawswelling), cartilage destruction, and bone resorption that occurs insemi-established type II collagen arthritis in female Lewis rats. Rats(10 per group) were injected intradermally (ID) with porcine type IIcollagen to induce arthritis; one group of rats (4 per group) served asvehicle-treated naïve controls. The animals were dosed PO, twice daily(BID) on study days 6 through (animals #1-5 in each group, animals #1-2in naïve group) or 21 (animals #6-10 in each group, animals #3-4 innaïve group) with vehicle (methylcellulose, viscosity: 1500 cP),Quercetin (230 mg/kg), Butyric acid (420 mg/kg), Compound 5 (100 mg/kgor 500 mg/kg), or Compound 45 (500 mg/kg). Positive controls weretreated PO, BID with the reference compound Tofacitinib citrate (10mg/kg). The rats were euthanized for necropsy on study day 21 (animals#1-5 in each group, animals #1-2 in naïve group) or 22 (animals #6-10 ineach group, animals #3-4 in naïve group). Efficacy evaluation was basedon animal body weights (Table 9), daily ankle caliper measurements (FIG.4), ankle diameter expressed as area under the curve (AUC) (Table 10),terminal hind paw weights (Table 11), histopathologic evaluation of hindpaws (Ankle Inflammation score: Table 12, Ankle Pannus score: Table 13,Ankle Cartilage Damage score: Table 14, Ankle Bone Resorption score:Table 15, Ankle Periosteal Bone score: Table 16, Ankle Periosteal BoneWidth: Table 17) and knees (Knee Inflammation score: Table 18, KneePannus score: Table 19, Knee Cartilage Damage score: Table 20, Knee BoneResorption score: Table 21), and relative spleen to body weight ratio(Table 22). Terminal serum was analyzed by enzyme-linked immunosorbentassay (ELISA) for anti-type II collagen antibodies (IgG) (Table 23).Treatment with compound 5 showed significant beneficial effects onspleen weights relative to body weight, ankle diameter AUC, ankle pannusscore, ankle bone resorption score, and ankle periosteal bone score. Allanimals survived to study termination. Statistics were conducted usingordinary one-way ANOVA with Dunnett's multiple comparison correction inGraphPad Prism.

The results of the study are summarized in Tables 9-23 below and FIG. 4.

TABLE 9 Change in body weight, day −1 to 21 p-value (if Mean bodysignificant), weight change, compared to Treatment day −1 to 21 (%)vehicle control Vehicle Control −12.5 Tofacitinib citrate (10 mg/kg)20.2 p < 0.0001 Quercetin (230 mg/kg) −13.9 Butyric acid (420 mg/kg)−2.3 p < 0.05  Compound 5 (100 mg/kg) −7.1 Compound 5 (500 mg/kg) −10.1Compound 45 (500 mg/kg) −6.3 Naïve Vehicle 17.3 p < 0.0001

TABLE 10 Ankle Diameter AUC p-value (if Mean Ankle significant),Diameter AUC compared to Treatment (mm*day) vehicle control VehicleControl 95.29 Tofacitinib citrate (10 mg/kg) 73.82 p < 0.0001 Quercetin(230 mg/kg) 96.85 Butyric acid (420 mg/kg) 92.46 Compound 5 (100 mg/kg)89.38 p < 0.05  Compound 5 (500 mg/kg) 93.57 Compound 45 (500 mg/kg)94.20 Naïve Vehicle 73.09 p < 0.0001

TABLE 11 Terminal hind paw weights p-value (if Mean Terminalsignificant), hind paw compared to Treatment weights (g) vehicle controlVehicle Control 2.4776 Tofacitinib citrate (10 mg/kg) 1.4321 p < 0.0001Quercetin (230 mg/kg) 2.5210 Butyric acid (420 mg/kg) 2.4759 Compound 5(100 mg/kg) 2.3990 Compound 5 (500 mg/kg) 2.3994 Compound 45 (500 mg/kg)2.3988 Naïve Vehicle 1.3854 p < 0.0001

TABLE 12 Ankle inflammation score, 0-5 (0 = normal, 5 = severe) p-value(if Mean Ankle significant), inflammation compared to Treatment score(0-5) vehicle control Vehicle Control 2.4776 Tofacitinib citrate (10mg/kg) 1.4321 p < 0.0001 Quercetin (230 mg/kg) 2.5210 Butyric acid (420mg/kg) 2.4759 Compound 5 (100 mg/kg) 2.3990 Compound 5 (500 mg/kg)2.3994 Compound 45 (500 mg/kg) 2.3988 Naïve Vehicle 1.3854 p < 0.0001

TABLE 13 Ankle Pannus score, 0-5 (0 = normal, 5 = severe) p-value (ifMean significant), Ankle Pannus compared to Treatment score (0-5)vehicle control Vehicle Control 3.45 Tofacitinib citrate (10 mg/kg) 0.00p < 0.0001 Quercetin (230 mg/kg) 3.55 Butyric acid (420 mg/kg) 2.95Compound 5 (100 mg/kg) 2.53 p < 0.05  Compound 5 (500 mg/kg) 3.33Compound 45 (500 mg/kg) 3.48 Naïve Vehicle 0.00 p < 0.0001

TABLE 14 Ankle Cartilage Damage score, 0-5 (0 = normal, 5 = severe) MeanAnkle p-value (if Cartilage significant), Damage score compared toTreatment (0-5) vehicle control Vehicle Control 4.43 Tofacitinib citrate(10 mg/kg) 0.00 p < 0.0001 Quercetin (230 mg/kg) 4.70 Butyric acid (420mg/kg) 3.80 Compound 5 (100 mg/kg) 3.55 Compound 5 (500 mg/kg) 4.45Compound 45 (500 mg/kg) 4.20 Naïve Vehicle 0.00 p < 0.0001

TABLE 15 Ankle Bone Resorption score, 0-5 (0 = normal, 5 = severe) Meanp-value (if Ankle Bone significant), Resorption compared to Treatmentscore (0-5) vehicle control Vehicle Control 3.45 Tofacitinib citrate (10mg/kg) 0.00 p < 0.0001 Quercetin (230 mg/kg) 3.55 Butyric acid (420mg/kg) 2.95 Compound 5 (100 mg/kg) 2.53 p < 0.05  Compound 5 (500 mg/kg)3.33 Compound 45 (500 mg/kg) 3.48 Naïve Vehicle 0.00 p < 0.0001

TABLE 16 Ankle Periosteal Bone score, 0-5 (0 = normal, 5 = severe) MeanAnkle p-value (if Periosteal significant), Bone score compared toTreatment (0-5) vehicle control Vehicle Control 3.85 Tofacitinib citrate(10 mg/kg) 0.00 p < 0.0001 Quercetin (230 mg/kg) 3.95 Butyric acid (420mg/kg) 3.08 Compound 5 (100 mg/kg) 2.68 p < 0.05  Compound 5 (500 mg/kg)3.70 Compound 45 (500 mg/kg) 3.38 Naïve Vehicle 0.00 p < 0.0001

TABLE 17 Ankle Periosteal Bone width (um) Mean Ankle p-value (ifPeriosteal significant), Bone width compared to Treatment (um) vehiclecontrol Vehicle Control 727.65 Tofacitinib citrate (10 mg/kg) 0.00 p <0.0001 Quercetin (230 mg/kg) 727.65 Butyric acid (420 mg/kg) 573.30Compound 5 (100 mg/kg) 510.30 Compound 5 (500 mg/kg) 699.30 Compound 45(500 mg/kg) 630.00 Naïve Vehicle 0.00 p < 0.0001

TABLE 18 Knee Inflammation score, 0-5 (0 = normal, 5 = severe) p-value(if Mean Knee significant), Inflammation compared to Treatment score(0-5) vehicle control Vehicle Control 5.35 Tofacitinib citrate (10mg/kg) 0.10 p < 0.0001 Quercetin (230 mg/kg) 5.48 Butyric acid (420mg/kg) 4.83 Compound 5 (100 mg/kg) 5.23 Compound 5 (500 mg/kg) 5.65Compound 45 (500 mg/kg) 5.25 Naïve Vehicle 0.00 p < 0.0001

TABLE 19 Knee Pannus score, 0-5 (0 = normal, 5 = severe) p-value (ifMean Knee significant), Pannus compared to Treatment score (0-5) vehiclecontrol Vehicle Control 2.60 Tofacitinib citrate (10 mg/kg) 0.00 p <0.0001 Quercetin (230 mg/kg) 3.18 Butyric acid (420 mg/kg) 2.43 Compound5 (100 mg/kg) 2.08 Compound 5 (500 mg/kg) 2.65 Compound 45 (500 mg/kg)2.48 Naïve Vehicle 0.00 p < 0.0001

TABLE 20 Knee Cartilage Damage score, 0-5 (0 = normal, 5 = severe) MeanKnee p-value (if Cartilage significant), Damage compared to Treatmentscore (0-5) vehicle control Vehicle Control 3.55 Tofacitinib citrate (10mg/kg) 0.00 p < 0.0001 Quercetin (230 mg/kg) 3.98 Butyric acid (420mg/kg) 3.28 Compound 5 (100 mg/kg) 2.93 Compound 5 (500 mg/kg) 4.00Compound 45 (500 mg/kg) 3.63 Naïve Vehicle 0.00 p < 0.0001

TABLE 21 Knee Bone Resorption score, 0-5 (0 = normal, 5 = severe)p-value (if Mean Knee significant), Bone Resorption compared toTreatment score (0-5) vehicle control Vehicle Control 2.60 Tofacitinibcitrate (10 mg/kg) 0.00 p < 0.0001 Quercetin (230 mg/kg) 3.15 Butyricacid (420 mg/kg) 2.43 Compound 5 (100 mg/kg) 2.08 Compound 5 (500 mg/kg)2.65 Compound 45 (500 mg/kg) 2.40 Naïve Vehicle 0.00 p < 0.0001

TABLE 22 Relative Spleen to Body Weight p-value (if Relativesignificant), Spleen to Body compared to Treatment Weight (Mean) vehiclecontrol Vehicle Control 0.2779 Tofacitinib citrate (10 mg/kg) 0.1796  p< 0.0001 Quercetin (230 mg/kg) 0.2539 Butyric acid (420 mg/kg) 0.2416 p< 0.01 Compound 5 (100 mg/kg) 0.2453 p < 0.05 Compound 5 (500 mg/kg)0.2534 Compound 45 (500 mg/kg) 0.2534 Naïve Vehicle 0.2345 p < 0.05

TABLE 23 Serum IgG ELISA p-value (if significant), Serum IgG compared toTreatment (ug/mL) vehicle control Vehicle Control 6569.33 Tofacitinibcitrate (10 mg/kg) 2524.70 p < 0.0001 Quercetin (230 mg/kg) 7014.93Butyric acid (420 mg/kg) 5824.57 Compound 5 (100 mg/kg) 5937.67 Compound5 (500 mg/kg) 5350.41 Compound 45 (500 mg/kg) 5431.64 Naïve Vehicle 1.19p < 0.05 

Other Embodiments

Various modifications and variations of the described invention will beapparent to those skilled in the art without departing from the scopeand spirit of the invention. Although the invention has been describedin connection with specific embodiments, it should be understood thatthe invention as claimed should not be unduly limited to such specificembodiments. Indeed, various modifications of the described modes forcarrying out the invention that are obvious to those skilled in the artare intended to be within the scope of the invention.

Other embodiments are in the claims.

What is claimed is:
 1. A method of modulating an autoimmunity marker ortreating an autoimmune disorder in a subject in need thereof, the methodcomprising administering to the subject an effective amount of anacylated active agent selected from the group consisting of an acylatedcatechin polyphenol, acylated carotenoid, acylated mesalamine, acylatedsugar, acylated shikimic acid, acylated ellagic acid, acylated ellagicacid analogue, and acylated hydroxy benzoic acid.
 2. The method of claim1, wherein the autoimmunity marker is for ulcerative colitis or Crohn'sdisease.
 3. The method of claim 1, wherein the autoimmune disorder isulcerative colitis or Crohn's disease.
 4. The method of claim 1, whereina CYP1A1 mRNA level, intestinal motility, CD4⁺CD25⁺ Treg cell count,short chain fatty acids level, mucus secretion is increased followingthe administration of the acylated active agent to the subject; orwherein abdominal pain, gastrointestinal inflammation, gastrointestinalpermeability, gastrointestinal bleeding, intestinal motility, orfrequency of bowel movements is reduced following the administration ofthe acylated active agent to the subject; or wherein an interleukin-8(IL-8) level, macrophage inflammatory protein 1α (MIP-1α) level,macrophage inflammatory protein 1β (MIP-1p) level, NFκB level, induciblenitric oxide synthase (iNOS) level, matrix metallopeptidase 9 (MMP9)level, interferon γ (IFNγ) level, interleukin-17 (IL17) level,intercellular adhesion molecule (ICAM) level, CXCL13 level,8-iso-prostaglandin F_(2α) (8-iso-PGF2α) level IgA level, calprotectinlevel, lipocalin-2 level, or indoxyl sulfate level is reduced followingthe administration of the acylated active agent to the subject; orwherein the T_(h)1 cell count is modulated following the administrationof the acylated active agent to the subject.
 5. The method of claim 1,wherein the acylated active agent is selected from the group consistingof an acylated catechin polyphenol, acylated mesalamine, acylated sugar,acylated shikimic acid, and acylated hydroxybenzoic acid.
 6. The methodof claim 1, wherein the method comprises administering the acylatedactive agent to the subject orally.
 7. The method of claim 1, whereinthe acylated active agent is cleaved to release at least one fatty acid,at least one ketone body, at least one pre-ketone body, or at least oneamino acid metabolite.
 8. The method of claim 1, wherein the acylatedactive agent comprises a group containing a fatty acid, and wherein thegroup containing a fatty acid is a monosaccharide, sugar alcohol, orsugar acid having one or more hydroxyl groups substituted with a fattyacid acyl.
 9. The method of claim 1, wherein the acylated active agentcomprises a group containing a fatty acid.
 10. The method of claim 9,wherein the fatty acid is a short chain fatty acid.
 11. The method ofclaim 10, wherein the short chain fatty acid is acetyl, propionyl, orbutyryl.
 12. The method of claim 1, wherein the acylated active agentcomprises a group containing an amino acid metabolite.
 13. The method ofclaim 1, wherein the acylated active agent is an acylated mesalamine.14. The method of claim 1, wherein the acylated active agent is anacylated catechin polyphenol, acylated sugar, acylated shikimic acid,acylated hydroxybenzoic acid, acylated ellagic acid, or acylated ellagicacid analogue.
 15. The method of claim 1, wherein the acylated activeagent comprises at least one ketone body acyl or at least one amino acidmetabolite acyl.
 16. An acylated mesalamine of formula (II):

or a pharmaceutically acceptable salt thereof, wherein R¹ is H, alkyl,acyl, a group containing a fatty acid, a group containing a ketone bodyor pre-ketone body, or a group containing an amino acid metabolite; R²is H, alkyl, a group containing a fatty acid, a group containing aketone body or pre-ketone body, or a group containing an amino acidmetabolite; and each R³ is independently H, alkyl, acyl, a groupcontaining a fatty acid, a group containing a ketone body or pre-ketonebody, or a group containing an amino acid metabolite; or both R² groupscombine to form:

provided that the acylated mesalamine comprises at least one groupcontaining a fatty acid, at least one group containing a ketone body orpre-ketone body, or at least one group containing an amino acidmetabolite.
 17. The acylated mesalamine of claim 16, wherein R¹ is agroup containing a fatty acid.
 18. The acylated mesalamine of claim 17,wherein the group containing a fatty acid is bonded through a glycosidicbond.
 19. The acylated mesalamine of claim 16, wherein the groupcontaining a fatty acid is a monosaccharide, sugar alcohol, or sugaracid having one or more hydroxyl groups substituted with a fatty acidacyl.
 20. The acylated mesalamine of claim 16, wherein the fatty acid isa short chain fatty acid.
 21. The acylated mesalamine of claim 20,wherein the short chain fatty acid is butyryl.
 22. The acylatedmesalamine of claim 16, wherein R² is H.
 23. The acylated mesalamine ofclaim 16, wherein each R³ is H.
 24. The acylated mesalamine of claim 16,wherein the acylated mesalamine is a compound of the followingstructure:

or a pharmaceutically acceptable salt thereof.
 25. The acylatedmesalamine of claim 24, wherein the compound is of the followingstructure:

or a pharmaceutically acceptable salt thereof.
 26. The acylatedmesalamine of claim 24, wherein the compound is of the followingstructure:

or a pharmaceutically acceptable salt thereof.
 27. The acylatedmesalamine of claim 24, wherein the compound is of the followingstructure:

or a pharmaceutically acceptable salt thereof.
 28. A compositioncomprising an excipient and the acylated mesalamine of claim
 16. 29. Anacylated shikimic acid of the following structure:

or a salt thereof, where each R¹ is independently H, acyl, alkyl, agroup containing a fatty acid, a group containing a ketone body orpre-ketone body, or a group containing an amino acid metabolite; and R²is H, alkyl, a group containing a fatty acid, a group containing aketone body or pre-ketone body, or a group containing an amino acidmetabolite; provided that the compound includes at least one groupcontaining a fatty acid, group containing a ketone body or pre-ketonebody, or group containing an amino acid metabolite.
 30. An acylatedcatechin polyphenol of formula (I):

or a pharmaceutically acceptable salt thereof, wherein

is a single carbon-carbon bond or double carbon-carbon bond; Q is —CH₂—or —C(O)—; each R¹ and each R³ is independently H, halogen, —OR^(A),phosphate, or sulfate; R² is H or —OR^(A); each R^(A) is independentlyH, optionally substituted alkyl, a monosaccharide, a sugar acid, a groupcontaining a fatty acid, a group containing a ketone body or pre-ketonebody, a group containing an amino acid metabolite, or benzoyl optionallysubstituted with 1, 2, 3, or 4 substituents independently selected fromthe group consisting of H, hydroxy, halogen, a group containing a fattyacid, a group containing a ketone body or pre-ketone body, a groupcontaining an amino acid metabolite, an optionally substituted alkyl, anoptionally substituted alkoxy, a monosaccharide, a sugar acid,phosphate, and sulfate; each of n and m is independently 0, 1, 2, 3, or4; provided that the compound of formula (I) comprises at least onegroup containing a fatty acid, group containing a ketone body orpre-ketone body, or group containing an amino acid metabolite; andprovided that at least one group containing a fatty acid, when present,is a monosaccharide having one, two, three, or four hydroxylssubstituted with fatty acid acyls.